Extended Data Fig. 3: Autophagy status is unchanged in animals expressing sid-1 and in non-neuronal tissues after neuronal knockdown of early-acting autophagy genes.
(a) Mean neuronal GFP::LGG-1 and GFP::LGG-1(G116A) punctae in day 1 wild-type (sid-1 + /+) (n = 28, 29) and sid-1(qt9) (sid-1-/-) animals (n = 27, 27) with or without rgef-1p::sid-1 transgene (rgef-1p::sid-1 (+)) (n = 26, 29). Error bars are s.d. over 3 independent experiments. Comparison between strains: LGG-1: ns P = 0.75, P = 0.97, G116A: ns P = 0.98, P > 0.99. Comparison of lipidated and unlipidated structures: ****P < 0.0001, by two-way ANOVA with Tukey’s multiple comparisons test. (b) Mean sqst-1p::sqst-1::gfp fluorescence intensity in head region of day 1 wild-type (sid-1 + /+), and sid-1(qt9) (sid-1(-/-)) animals with or without rgef-1p::sid-1 transgene (rgef-1p::sid-1 (+)) on day 1 of adulthood. Error bars are s.d. of n = 31 animals over 3 independent experiments. ns P = 0.18 and P = 0.59 by one-way ANOVA with Dunnett’s multiple comparisons test. (c) GFP::LGG-1 punctae in intestinal cells of day 1 wild-type (sid-1 + /+) (n = 59) and sid-1(qt9) (sid-1-/-) animals (n = 62) with or without rgef-1p::sid-1 transgene (rgef-1p::sid-1 (+)) (n = 62). Violin plots with solid line indicating median and dashed lines indicating quartiles. ns P = 0.79, P = 0.99 by one-way ANOVA with Dunnett’s multiple comparisons test. (d) GFP::LGG-1 punctae in body-wall muscle areas of day 1 wild-type (sid-1 + /+) (n = 48) and sid-1(qt9) (sid-1-/-) animals (n = 45) with or without rgef-1p::sid-1 transgene (rgef-1p::sid-1 (+)) (n = 52). Violin plots with solid line indicating median and dashed lines indicating quartiles. ns P = 0.64, P = 0.70 by one-way ANOVA with Dunnett’s multiple comparisons test. (e) GFP::LGG-1 punctae in intestinal cells of day 1 sid-1; rgef-1p::sid-1 + rgef-1p::gfp; lgg-1p::gfp::lgg-1 animals after whole-life atg-7 (n = 65), or lgg-1/ATG8 (n = 48) RNAi compared to control (CTRL) (n = 81). Violin plots with solid line indicating median and dashed lines indicating quartiles. ns P = 0.98, P = 0.71 by one-way ANOVA with Dunnett’s multiple comparisons test. (f) GFP::LGG-1 punctae in body-wall muscle areas of day 1 sid-1; rgef-1p::sid-1 + rgef-1p::gfp; lgg-1p::gfp::lgg-1 animals after whole-life atg-7 (n = 54), or lgg-1/ATG8 (n = 53) RNAi compared to control (CTRL) (n = 49). Violin plots with solid line indicating median and dashed lines indicating quartiles. ns P = 0.34, P = 0.96 by one-way ANOVA with Dunnett’s multiple comparisons test.
