Figure 1
PUFA-deficient mutants fat-3 and daf-7;fat-3 exhibit a dauer-like or a Daf-c phenotype, respectively. (a) Unlike wild-type worms, fat-3(ok1126) mutants are incapable of synthesizing PUFAs and undergo a dauer-like arrest in the first generation when grown in cholesterol-free media. One-way analysis of variance p < 0.001. All pairwise multiple comparison procedures (Holm-Sidak method), (*) indicates statistically significant difference with fat-3/solvent, p < 0.001. Bars represent mean values and error bars represent standard errors. The number of independent experiments is n = 3 for all conditions. (b) While daf-7 worms form ~10% dauers at 20 ºC, fat-3 worms do not form dauers. In stark contrast, double mutant daf-7;fat-3 forms ~85% dauers at 20 ºC, and addition of DA suppresses dauer formation almost completely. One-way analysis of variance p < 0.001. All pairwise multiple comparison procedures (Holm-Sidak method): (*) indicates statistically significant difference with daf-7, p < 0.001. Bars represent mean values and error bars represent standard errors. The number of independent experiments is n = 5 for daf-7, and n = 13 for fat-3 and daf-7; fat-3. [DA] = 90 nM. (c) Introducing the fat-3 null-mutation into background daf-7 increases dauer formation significantly (dauers are indicated with arrowheads). Representative images are taken from at least three experiments. Scale bars, 0.5 mm. (d) daf-7;fat-3 dauers can partly bypass dauer arrest if grown in excess cholesterol (130 µM). (*) indicates statistically significant difference to control plates (13 µM cholesterol). t-test, p < 0.001. Bars represent mean values and error bars represent standard errors. The number of independent experiments is n = 14 for daf-7;fat-3 at 13 µM cholesterol, and n = 2 for daf-7;fat-3 at 130 µM cholesterol.
