(A) Schematic depiction of the oxygen-dependent fat loss assay. Worms were washed off food, and then fasted at either 21% (high) oxygen or 10% (low) oxygen. At the end of the fasting period, worms were fixed and stained with Oil Red O. (B) Worms of the indicated genotypes were subjected to the oxygen-dependent fat loss assay described in (A). White bars, fed; gray bars, fasted at 21% oxygen and black bars, fasted at 10% oxygen. Fat content was quantified for each genotype and condition. Data are expressed as a percentage of body fat in wild-type fed controls + SEM (n = 16–20). NS, not significant and *, p<0.05 by one-way ANOVA. (C) Worms of the indicated genotypes were subjected to the oxygen-dependent fat loss assay. Fat content was quantified for each genotype and condition. Data are expressed as a percentage of body fat in wild-type fed controls + SEM (n = 18–20). NS, not significant and *, p<0.05 by one-way ANOVA. (D) Oxygen consumption rate (OCR) in wild-type, gcy-33, gcy-36, and gcy-33;gcy-36 mutants. Data are presented as pmol/min/worm + SEM (n = 5–15). *, p<0.05 by two-way ANOVA.