Nutrient deprivation induces autophagy at region 2 within the germarium and in dying midstage egg chambers. (A) GFP-LC3 proteins were expressed in nurse cells (NC) but not in follicle cells (FC) by using the UASp/nanos Gal4 system. DAPI staining of nuclei is shown in blue. (B) UASp-GFP-LC3; nanos GAL4 flies were conditioned on yeast paste and had a diffuse GFP-LC3 pattern. Numerous GFP-LC3 puncta (green) at region 2 within germarium were observed in nutrient-deprived flies. Ovaries were stained with LTR in w¹¹¹⁸ flies. Germarium of nutrient-deprived w¹¹¹⁸ flies had an increase in punctate LTR staining (red) compared with well-fed germarium. (C) Degenerating stage 8 egg chambers (arrows) had numerous GFP-LC3 puncta (green) and an increase in LTR-positive dots (red) compared with healthy egg chambers (arrowheads). DAPI (blue) staining of nuclei is shown in the two panels on the right. (D) Degenerating stage 8 egg chambers (arrows) of nutrient-deprived Atg7 mutants (Atg7^d77/Atg7^d14) showed a dramatic decrease in LTR staining. DAPI staining of nuclei is shown in blue. Bars: (A and B) 20 μm; (C and D) 50 μm.