Worms deficient in <i>lipl-2</i>, an HLH-30 target gene, displayed starvation–refeeding mortality that was rescued by CeMM.

(A) lipl-2 mRNA abundance in au with values normalized to the control gene ama-1 determined by qPCR in L1 stage WT and hlh-30(lf) animals in the fed state (fed), after starvation for 33 hours (starved), and after starvation for 33 hours followed by refeeding on E. coli OP50 for 15 hours (OP50). N = 3 biological replicates/group. *P Fig 1A for lipl-2(lf) mutant animals and WT controls. *P lipl-2(lf) worms were analyzed after 10 days of starvation and 0 or 15 hours of complete CeMM exposure, followed by 48 hours on E. coli OP50. *P hlh-30(lf) worms with lipl-2 overexpression (lipl-2(oe);hlh-30(lf)) and hlh-30(lf) (as controls) analyzed after 33 hours of starvation followed by exposure for 15 hours to CeMM minimal solution (lacking glucose, β-sitosterol in Tween 80, amino acids, and nucleic acids) supplemented with β-sitosterol in Tween 80 (lipids) or glucose, followed by 48 hours on E. coli OP50. Similarly modeled worms without exposure to CeMM are shown as control. *P N = 3 biological replicates/group with 50 worms/time point for panels B–D. Bars and values indicate mean ± SEM. Raw data for A–E are in S1 Data. ama-1, amanitin-binding subunit of RNA polymerase II; au, arbitrary unit; CeMM, C. elegans maintenance medium; hlh-30, basic helix–loop–helix transcription factor 30; hlh-30(lf), loss-of-function tm1978 mutation hlh-30; lipl-2, lysosomal lipase 2; lipl-2(lf), loss-of-function mutation lipl-2; lipl-2(oe);hlh-30(lf), hlh-30(lf) with overexpressed lipl-2; L1, first larval stage; qPCR, quantitative PCR; SEM, standard error of the mean; WT, wild type.