NASC

N2105605

Name: P1Y, mCITRINE-1xFYVE(HRS)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with mCITRINE (YFP), localized in late endosome and tonoplast

N2105606

Name: P3Y, 1xPX(p40)-mCITRINE

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with mCITRINE (YFP), localized in late endosome and tonoplast

N2105607

Name: P5Y, mCITRINE-1xPH(FAPP1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with mCITRINE (YFP), localized at the plasma membrane, post-golgi/endosomal compartment (early endosome/TGN + recycling endosome) and weakly in the golgi

N2105608

Name: P7Y, mCITRINE-1xPH(OSBP)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with mCITRINE (YFP), localized at the plasma membrane

N2105609

Name: P14Y, mCITRINE-1xPH(PLC)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4,5-phosphate) fused with mCITRINE (YFP), localized in the cytosol and at the plasma membrane

N2105610

Name: P15Y, mCITRINE-1xTUBBY-C

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4,5-phosphate) fused with mCITRINE (YFP), localized at the plasma membrane and in the nucleus

N2105611

Name: P18Y, mCITRINE-2xFYVE(HRS)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with mCITRINE (YFP), localized in late endosome and in the tonoplast in some cells

N2105612

Name: P21Y, mCITRINE-2xPH(FAPP1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with mCITRINE (YFP), localized at the plasma membrane, post-golgi/endosomal compartment (early endosome/TGN + recycling endosome) and weakly in the golgi

N2105613

Name: P24Y, mCITRINE-2xPH(PLC)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with mCITRINE (YFP), localized at the plasma membrane

N2105614

Name: P1R, 2xmCHERRY-1xFYVE(HRS)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized in the cytosol and late endosome

N2105615

Name: P3R, 1xPX(p40)-2xmCHERRY

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized in late endosome and tonoplast

N2105616

Name: P5R, 2xmCHERRY-1xPH(FAPP1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized at the plasma membrane, post-golgi/endosomal compartment (early endosome/TGN + recycling endosome) and weakly in the golgi

N2105617

Name: P7R, 2xmCHERRY-1xPH(OSBP)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized at the plasma membrane

N2105618

Name: P14R, 2xmCHERRY-1xPH(PLC)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4,5-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized in the cytosol and at the plasma membrane

N2105619

Name: P15R, 2xmCHERRY-1xTUBBY-C

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4,5-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized at the plasma membrane and in the nucleus

N2105620

Name: P18R, 2xmCHERRY-2xFYVE(HRS)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized in late endosome and in the tonoplast in some cells

N2105621

Name: P21R, 2xmCHERRY-2xPH(FAPP1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized at the plasma membrane, post-golgi/endosomal compartment (early endosome/TGN + recycling endosome) and weakly in the golgi

N2105622

Name: P24R, 2xmCHERRY-2xPH(PLC)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4-phosphate) fused with 2xmCHERRY (Red fluorescent protein), localized at the plasma membrane

N2105623

Name: P3C, 2xCyPET-1xPX(p40)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with 2xCyPET (Cyan fluorescent protein), localized in late endosome and tonoplast

N2105624

Name: P5C, 1xPX(p40)-UBQ10PROM::2XCypET-1XPH(FAPP1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI4P (phosphatidylinositol 4-phosphate) fused with 2xCyPET (Cyan fluorescent protein), localized at the plasma membrane, post-golgi/endosomal compartment (early endosome/TGN + recycling endosome) and weakly in the golgi

N2105625

Name: P14C, UBQ10prom::2xCyPet-1PH(PLCd1)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI(4,5)P2 (phosphatidylinositol 4,5-phosphate) fused with 2xCyPET (Cyan fluorescent protein), localized in the cytosol and at the plasma membrane

N2105626

Name: P18C, UBQ10prom::2xCyPet-2xFYVE(HRS)

Price: £11.00

Donor Ecole Normale Supérieure de Lyon Yvon Jaillais

Stock type: individual line

Material type: seed

Description Each PIPline construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants were transformed by dipping. Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for CITRINE-tagged PIPline, hygromycin for CHERRY-tagged PIPline and kanamycin for CyPET-tagged PIPline). 24 independent T1s were selected for each PIPline. In the T2 generation at least 3 independent transgenic lines were selected using the following criteria when possible: i) good expression level in the root for detection by confocal microscopy, ii) uniform expression pattern, iii) single insertion line (1 sensitive to 3 resistant segregation ratio) and, iv) line with no obvious abnormal developmental phenotypes. PIPlines for which we could not find fluorescence out of 24 independent lines or that did not associated with any membrane compartments (including the PM) were not further analysed. The remaining PIPlines were rescreened in T3 using similar criteria as in T2 with the exception that we selected homozygous lines (100% resistant). At this step, we selected one transgenic line for each PIPline that was used for further analysis and crosses and that was distributed to the Arabidopsis stock centres.

Phenotype geneticaly encoded biosensor for PI3P (phosphatidylinositol 3-phosphate) fused with 2xCyPET (Cyan fluorescent protein), localized in late endosome and in the tonoplast in some cells