Fig. 3: scl-12&13 mutations lead to cholesterol-dependent impaired dauer exit.

a Percentage of daf-2 and scl-12&13;daf-2 populations that develop into L4 larvae 48 h after dauer recovery was induced by temperature switch to 15 °C, control (ethanol; black) and when treated with 200 nm (25S)-Δ7-dafachronic acid (DA; red). Average ± SD of four independent experiments with a minimum of 15 worms, two-way ANOVA with post hoc Sidak’s multiple comparisons test comparing daf-2 Ctrl versus scl-12&13;daf-2 Ctrl showed p < 0.0001 and scl-12&13;daf-2 Ctrl versus scl-12&13;daf-2 DA p < 0.0001. b Growth/length in mm of daf-2 and scl-12&13;daf-2 after induction of dauer exit by temperature switch and the effect of DA thereon. Individual values of four independent experiments with a minimum of 10 worms, two-way ANOVA comparing daf-2 Ctrl versus scl-12&13;daf-2 Ctrl showed p = 0.0001, and scl-12&13;daf-2 Ctrl versus scl-12&13;daf-2 DA p < 0.0001. c Scheme for the experimental procedure in (d). d Percentage of daf-2 (black) and scl-12&13;daf-2 (gray) populations that develop into L4 larvae 48 h after dauer recovery was induced by temperature switch to 15 °C. Worms were grown with or without cholesterol (+ or -) from egg to dauer (“Entry”) and with or without cholesterol from dauer to L4 while exiting dauer (“Exit”). Average ± SD and individual values of four independent experiments with a minimum of 15 worms, two-way ANOVA with post hoc Sidak’s multiple comparisons tests comparing daf-2 chol+/chol+ versus scl-12&13;daf-2 chol+/chol+ showed p < 0.0001, daf-2 chol+/chol− versus scl-12&13;daf-2 chol+/chol− p = 0.0002, daf-2 chol-/chol+ versus scl-12&13;daf-2 chol-/chol+ p = 0.0432 as well as daf-2 chol+/chol+ versus daf-2 chol−/chol+ p = 0.0004 and daf-2 chol+/chol+ versus daf-2 chol−/chol− p = 0.0001.