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. 2014 May 26;205(4):477–492. doi: 10.1083/jcb.201303144

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© 2014 DeVorkin et al.

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Figure 3. — Ref(2)P analyses confirm that Dcp-1 is required for autophagic flux in ovaries. Staining shows DNA, Ref(2)P, and Armadillo. Zoomed insets show Ref(2)P staining. (A) Nondegenerating midstage egg chambers from fed w¹¹¹⁸ flies showed Ref(2)P staining in follicle cells and nurse cells. (B) Starved w¹¹¹⁸ flies contained degenerating midstage egg chambers with reduced Ref(2)P. (C) A representative Western blot of ovaries from w¹¹¹⁸ and Atg7^d77/d14 flies subjected to fed or starvation conditions for 4 d. Ref(2)P was detected by immunoblotting. Actin served as a loading control. Densitometry was performed to quantitate Ref(2)P protein levels relative to actin. Graph represents ± SD from five independent experiments (n = 5). Statistical significance was determined using a two-tailed Student’s t test. *, P = 0.004 for fed samples; *, P = 0.008 for starved samples. (D) Starved Atg7 flies showed an accumulation of Ref(2)P in the follicle cells and nurse cells of degenerating midstage egg chambers. (E) Starved Dcp-1^Prev1 flies contained increased levels of Ref(2)P in degenerating midstage egg chambers. (F) A representative Western blot of ovaries from w¹¹¹⁸ and Dcp-1^Prev1 flies that were subjected to fed or starvation conditions for 4 d. Ref(2)P was detected by immunoblotting. Actin served as a loading control. Densitometry was performed to quantitate Ref(2)P protein levels relative to actin. Graph represents ± SD from five independent experiments (n = 5). Statistical significance was determined using a two-tailed Student’s t test. *, P = 0.03. Bars: (main images) 25 µm; (zoomed images) 10 µm.