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Figure 6.
Pex30 and PA distribution is affected by PC levels in sei1Δ cells. (A) Yeast phospholipid synthesis pathway. The genes targeted in the screen are indicated in red. (B) WF images of the indicated strains endogenously expressing Pex30-2xmCherry. Cells were grown in synthetic media with or without 2 mM choline supplementation until logarithmic phase. Yellow arrowheads denote Pex30-2xmCherry accumulation. Bar = 4 μm. (C) Quantification of experiment in B showing percent cells with Pex30-2xmCherry accumulation. Bars show mean from three independent experiments and SEM. 100 cells from each replicate were analyzed and compared using two-way ANOVA and Tukey’s multiple comparison test. Means were compared against sei1Δ. Additional comparisons with choline treatment are shown and are not significant (*P < 0.05, ****P < 0.001). (D) WF images of sei1opi3Δ cells in logarithmic phase endogenously expressing Pex30-2xmCherry and OPI3 on a plasmid. Yellow arrows denote Pex30-2xmCherry accumulation. The graph on the right is quantification of Pex30-2xmCherry accumulation in sei1opi3Δ with or without overexpression of OPI3. Bars show mean from two independent experiments. 100 cells from each replicate were analyzed. Bar = 4 μm. (E) WF images of the indicated strains expressing Opi1-GFP on a plasmid. Cells were grown in synthetic media with or without 2 mM choline supplementation. White arrowheads denote Opi1-GFP accumulation. Bar = 4 μm. (F) Quantification of experiment in E showing percent cells with Opi1-GFP puncta. Bars show mean from three independent experiments and SEM. 100 cells per genotype from each replicate were analyzed and compared using two-way ANOVA and Tukey’s multiple comparison test (*P < 0.05 and **P < 0.01). WF, widefield images.

Pex30 and PA distribution is affected by PC levels in sei1Δ cells. (A) Yeast phospholipid synthesis pathway. The genes targeted in the screen are indicated in red. (B) WF images of the indicated strains endogenously expressing Pex30-2xmCherry. Cells were grown in synthetic media with or without 2 mM choline supplementation until logarithmic phase. Yellow arrowheads denote Pex30-2xmCherry accumulation. Bar = 4 μm. (C) Quantification of experiment in B showing percent cells with Pex30-2xmCherry accumulation. Bars show mean from three independent experiments and SEM. 100 cells from each replicate were analyzed and compared using two-way ANOVA and Tukey’s multiple comparison test. Means were compared against sei1Δ. Additional comparisons with choline treatment are shown and are not significant (*P < 0.05, ****P < 0.001). (D) WF images of sei1opi3Δ cells in logarithmic phase endogenously expressing Pex30-2xmCherry and OPI3 on a plasmid. Yellow arrows denote Pex30-2xmCherry accumulation. The graph on the right is quantification of Pex30-2xmCherry accumulation in sei1opi3Δ with or without overexpression of OPI3. Bars show mean from two independent experiments. 100 cells from each replicate were analyzed. Bar = 4 μm. (E) WF images of the indicated strains expressing Opi1-GFP on a plasmid. Cells were grown in synthetic media with or without 2 mM choline supplementation. White arrowheads denote Opi1-GFP accumulation. Bar = 4 μm. (F) Quantification of experiment in E showing percent cells with Opi1-GFP puncta. Bars show mean from three independent experiments and SEM. 100 cells per genotype from each replicate were analyzed and compared using two-way ANOVA and Tukey’s multiple comparison test (*P < 0.05 and **P < 0.01). WF, widefield images.

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