Fig. 6

Key components of the autophagy pathway, other than dAtg1, do not modify the ey > hid-p35 phenotype. (a) Results of the suppression of ey > hid-p35 using RNAi targeting components of the autophagy pathway in Drosophila. Representative RNAi results for each gene were shown. Compared to the control where no RNAi was used, knockdown of dAtg1 significantly increases the percentage of weak phenotype or wildtype-like ey > hid-p35 flies to about 80 %. However, knockdown of dAtg3, dAtg6, dAtg8a, dAtg8b, dAtg9, dAtg17, vps15, and vps34 does not suppress the ey > hid-p35 overgrowth phenotype. In contrast, expression of a kinase dead form of TOR (TOR^TED) or knockdown of raptor, both of which cause activation of dAtg1, enhances the AiP phenotype. However, RNAi targeting unc-76, which mediates the function of dAtg1 in neuronal development, does not suppress the overgrowth of ey > hid-p35 flies. Room temperature (RT) was used in some cases due to strong lethality caused by expressing these RNAi lines at 25 °C in the background of ey > hid-p35. The ey > hid-p35 flies display comparable overgrowth phenotypes at 25 °C and RT. (b–c’) Late third instar eye discs labeled with cCasp3, Wg, and ELAV. Neither dAtg13 null mutants (dAtg13 ^Δ74) (b, b’) nor dAtg7 null mutants (dAtg7 ^Δ14/ dAtg7 ^Δ77) (c, c’) inhibit overgrowth, cCasp3 labeling and ectopic Wg expression (b’, b’; arrows) in ey > hid-p35 discs (at least 40 discs were analyzed for each genotype)