S1 Fig -

Throughout this figure: Scale bars = 200μm, Error bars = S.E.M. N = number of independent biological replicates. Statistical significance was assessed via ratio t-test, *p≤ 0.05, **p≤0.01, ***p≤0.001 and ****p≤0.0001. (A) Overview of the workflow of the screen for genes altering fat storage in C. elegans. (B) Representative images of the body fat content and distribution in WT(N2) and rrf-3 RNAi sensitive mutant worms. N = >5. (C) Representative images of the body fat content and distribution as made evident with ORO in worms treated with RNAi against the obesity candidate genes from the L1 stage. Asterisks denote those RNAi treatments that led to developmental delay. (D) Representative images of the age-dependent increases in fat stores as revealed by staining L3, L4, and 1-day adult worms with ORO. N = >5. (E) Retesting of ORO staining in worms treated with hsp-4 RNAi from the L3 stage showed no effect of hsp-4 knockdown on C. elegans body fat content. (F) ORO quantification in worms treated with hsp-4 RNAi as represented in panel E. Each data point represents the ORO intensity in one worm. Individual intensity values were normalized to the mean value of the EV RNAi control. N = 3.

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