FIGURE 6:

Stage-specific nutrient deprivation reduces PVD ERES number and somatodendritic size. (A) Confocal-fluorescence microscopy closeups of PVD soma and endogenous ERES marker, GFP::SEC-16, 1 d after PVD birth in fed or 1-d starved WT animals (scale bar = 5 µm). PVDs in fed and starved animals were quantified for (B) number of PVD GFP::SEC-16 puncta (n = 12 per condition), and (C) soma size (n = 12 per condition) measured as described in Methods. Significance determined by Student’s t test two-tailed, ****p < 0.0001. (D) Number of GFP::SEC-16 puncta in PVD soma of fed vs. starved animals in Cre-control vs let-363-floxed (left, n ≥ 8 animals for each condition) and WT vs. mec-3 animals (right, n ≥ 10 animals for each condition). Animals were counted 1 d after PVD birth except where indicated. Fed animals at this time point were in late L3 or early L4 stages. (E) Quantification of PVD quaternary dendrite number vs. ERES number in either animals that were removed from food 7 h before PVD birth and starved for 1 d (pink, n = 9), animals that were removed from food just after PVD birth and starved for 1 d (green, n = 9), or animals that were fed for the duration of the experiment until 1 d after PVD birth at the L4 larval stage (black, n = 7). Points shown are mean values for each condition with standard deviations indicated by error bars. Pearson correlation between quaternary dendrite number and ERES number across conditions: r = 0.7122, R2 = 0.5072, p (two-tailed) < 0.0001. Color coding corresponds to diagram of starvation timelines in Supplemental Figure S3. (F) Number of GFP::SEC-16 puncta in WT animals removed from nutrients at different stages of PVD development, and starved for either 1 d and recovered (left, n ≥ 11), or 2 d (middle and right, n ≥ 9). Significance for (D) and (E) determined by one-way ANOVA, *p < 0.05,**p < 0.01,***p < 0.001, ****p < 0.0001. Results were additionally repeated in at least two independent experiments.