CeMM rescued impaired survival in starved <i>hlh-30(lf)</i> worms.

(A) Schematic depicting the starvation/refeeding protocol with the addition of a variable period of CeMM exposure after starvation in M9 and before transfer to NGM dishes with E. coli OP50 (see Fig 1A legend for details). (B) WT and hlh-30(lf) worms were analyzed after 33 hours of starvation and variable periods of CeMM exposure followed by 48 hours on E. coli OP50 as “Alive after Refeeding.” N = 3 biological replicates with 50 worms/time point for all experiments. Bars indicate mean ± SEM. *P y-axis) and refed with E. coli OP50 or CeMM and analyzed for “Alive after Refeeding,” as described in in the legend for Fig 1A. N = 3 biological replicates with approximately 50 worms/time. Values indicate mean ± SEM. *P hlh-30(lf) worms were analyzed after 33 hours of starvation and 15 hours of exposure to modified formulations of CeMM followed by 48 hours on E. coli OP50 as “Alive after Refeeding.” For removal experiments (panel D), otherwise complete CeMM was prepared lacking heme, AAs, β-sito in Tween 80, Glu, or NAs. For addition experiments (panel E), CeMM minimal solution lacking Glu, β-sito in Tween 80, AAs, and NAs was supplemented with individual nutrients. N = 3 biological replicates with 50 worms/time point for all experiments. Bars indicate mean ± SEM. *P hlh-30(lf) worms were analyzed for “Alive after Refeeding” after 33 hours of starvation and 48 hours of refeeding with standard E. coli OP50 or E. coli OP50 supplemented with glucose and Tween 80. Bars indicate mean ± SEM. N = 3 biological replicates/group with approximately 50 worms/condition. *P S1 Data. AA, amino acid; CeMM, C. elegans maintenance medium; Glu, glucose; hlh-30, basic helix–loop–helix transcription factor 30; hlh-30(lf), loss-of-function tm1978 mutation hlh-30; NA, nucleic acid; NGM, nematode growth medium; SEM, standard error of the mean; WT, wild type; β-sito, β-sitosterol.