Autophagy is important in certain FC subtypes for DA formation. Spatially restricted GAL4 driver lines were used to express ATG1, ATG4, ATG5 and ATG8 RNAi and the effect of autophagy knock down on the dorsal appendage (DA) phenotype was quantified. Downregulation of ATG gene expression with the broad e22c-GAL4 driver causes the most severe DA defects (A and C-C”). Knocking down ATG genes with slbo-GAL4 and fru-GAL4 causes similar strong phenotypes (A and D-E’). Overexpression of ATG RNAi with c355-GAL4, c306-GAL4, 109-30-GAL4 and upd-GAL4 only generated a low percentage of eggs with defective DAs (A and F, F’). Similarly, the controls (UAS>lacZ, ey-GAL4) only occasionally showed defective eggs (A and B). A: Error bars show S.D. of the mean, P-values: * P<0.05, ** P<0.01, *** P<0.001. Eggs were collected from 5 females for each genotype, n=3. B-F’: Anterior is to the top, posterior to the bottom, dorsal to the front. Scale bar: 100 μm. Genotypes: GAL4 driver see Figure 2, ATG-RNAi lines see Additional file 2: Figure S2.