Fig. S5

Fig. S5 Up-regulated autophagy is not a shared feature of all zebrafish intestinal mutants. (A) Western blot analysis of LC3 in protein extracts of WT, setebos (set^s453) and caliban (clbn^s846) larvae. Actin was used as a loading control. (B) The levels of LC3II were quantitated by densitometric analysis of three independent Western blots. Chloroquine-treated set^s453 larvae at 96 hpf contain significantly higher LC3II levels compared to their chloroquine-treated WT siblings; meanwhile, LC3II levels are similar in chloroquine-treated set^s453 larvae and WT larvae treated with rapamycin and chloroquine. There are no significant differences between LC3II levels in clbn^s846 larvae and their WT siblings at 120 hpf, in the presence and absence of chloroquine. Data are represented as mean +/ SD (n = 3), *p<0.05. (C–H) Transmission electron micrographs of transverse sections of WT (C, E, G) and clbn^s846 larvae (D, F, H) through the intestinal bulb region at 120 hpf. There are negligible numbers of autophagosomes/autolysosomes in the IECs of WT and clbn^s846 larvae. Scale bars = 50 μm (C, D); 10 μm (E, F); 5 μm (G–H). ib, intestinal bulb; n, nucleus; m, mitochondria; mv, microvilli.