Figure 4.

Pex30/Pex29 complex contributes to spatial organization of LDs at the NVJ. (A) Localization of Pex30 and LDs in cells with the indicated genotype during early stationary phase. Pex30 was endogenously tagged with mNG (Pex30-mNG) and LDs were stained with the neutral lipid dye MDH. Bar, 5 µm. (B) Time-course analysis of Pex30-mNG distribution in WT and pex29Δ cells. Exponentially growing cells were monitored through diauxic shift into early stationary phase as indicated. The formation of NVJ was monitored by endogenous Nvj1-tdTomato, and LDs were stained with the neutral lipid dye MDH. Yellow and red arrowheads indicate cells with and without NVJ-clustered LDs, respectively. (C) Quantification of cells with the indicated genotype displaying LDs clustered at the NVJ during diauxic shift (7.5 h) and stationary phase (24 h). Three independent experiments were analyzed (>100 cells/genotype/experiment were counted). Bars represent SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were done to compare the percentage of cells with the WT condition (**, P < 0.01; *, P < 0.05). (D) Quantification of cells with the indicated genotype displaying Nvj1 at the NVJ during diauxic shift (7.5 h) and stationary phase (24 h). Three independent experiments were analyzed (>100 cells/genotype/experiment were counted). Bars represent SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were done to compare the percentage of cells with the WT condition (**, P < 0.01). (E) Quantification of cells with the indicated genotype displaying clustered LDs during early stationary phase. LDs were classified as clustered or nonclustered depending on their distribution, as depicted in the cartoon (on the right). Three independent experiments were analyzed (>100 cells/genotype/experiment were counted). Bars represent SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were done to compare the percentage of cells with LD clusters with the WT condition (****, P < 0.0001; ns, P > 0.5).