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. 2015 Sep 14;11(8):1358–1372. doi: 10.1080/15548627.2015.1063766

Figure 7.

Figure 7.

Gyf is essential for developmental, starvation-induced and physiological autophagy. (A–H) Fat bodies (FB) of wandering-stage third instar larvae (A–D) or feeding-stage third instar larvae that were placed on 20% sucrose solution for 3 h (E–H) of WT and GyfMI/MI flies were subjected to anti-Atg8a immunostaining (A, C, E and G) or LysoTracker Red (Lys) staining (B, D, F and H). Hoechst 33258 (DNA) was used to visualize nuclei. Scale bars: 50 μm. Number of Atg8a or LysoTracker Red puncta per cell in fat bodies are presented as means ±standard error (n ≥ 40) (C, D, G and H). (I–L) Heads (I and J) and thoraxes (K and L) of 1-wk-old WT and GyfMI/MI flies were analyzed through immunoblotting of Atg8a-I (16 kDa), Atg8a-II (14 kDa) and Actin (40 kDa). The ratios of Atg8a-II to Atg8a-I are presented as means ± standard error (n = 7) (J and L). (M and N) Thoraxes of 1-wk-old WT and GyfMI/MI flies were analyzed through immunoblotting of phospho-Thr398 S6k (70 kDa), phospho-Ser505 Akt1 (60 and 70 kDa), Atg1 (120 kDa) and Actin (40 kDa). The immunoblot results were quantified and presented as means ± standard error (n = 4). P values were calculated using the Student t-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, not significant.