Extended Data Fig. 4: Effects of H3K4me3 modifier or hsf-1 depletion and oleic acid supplementation on lipid ontology enrichment and lipid droplet formation (related to Fig. 4).

(a) Mass spectrometry analysis of lipid levels in Q40::YFP animals maintained on L4440, ash-2, hsf-1, or ash-2 + hsf-1 RNAi was conducted, and significantly altered lipids between indicated conditions were analyzed for ontology enrichment using LION/web analysis. Enriched categories in lipids that increased in each comparison are shown in red, and lipids that decreased in each comparison shown in blue, with statistically significant categories being color-coded by –log(FDR q-value). (b) Changes in saturated, monounsaturated, and polyunsaturated fatty acids in phospholipids in day 1 adult Q40::YFP worms maintained on L4440, ash-2, hsf-1, ash-2 + hsf-1 in the absence or presence of oleic acid (0.8 mM). (c, d) LIU1 (dhs-3p::dhs-3::GFP) lipid droplet reporter animals were maintained on control (L4440), ash-2, hsf-1, or ash-2 + hsf-1 RNAi, and fluorescence was imaged at day 1 (c) and day 5 (d) of adulthood. Representative images are shown. Scale bar is 0.1 mm. (e) Quantification of the dhs-3p::dhs-3::GFP fluorescence in (d). (f) mRNA levels of hsp-16.2 and hsf-1 in day 10 adult Q40::YFP worms maintained on L4440 or ash-2 RNAi. (g) Fluorescence of day 10 adult LIU1 (dhs-3p::dhs-3::GFP) animals maintained on L4440 or ash-2 RNAi. Fluorescence is quantified in (h). At least 20 animals assessed per condition. Results are representative of at least three independent experiments. Error bars represent SEM. Statistical significance was assessed by one-way ANOVA (e), one sample, two-tailed t-test (f) or Kruskal-Wallis test (h); ***, p < 0.001; ****, p < 0.0001.