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. 2013 May 13;201(4):531–539. doi: 10.1083/jcb.201211160

Figure 1.

Figure 1.

Syx17, usnp, and VAMP7 are required for autophagy in Drosophila. (A–C) Syx17 (A), usnp (B), or VAMP7 (C) depletion in GFP-marked fat cell clones leads to formation of numerous small, mostly perinuclear mCherry-Atg8a dots, unlike the larger, brighter, evenly distributed punctae in surrounding control cells of starved larvae. (D–F) Knockdown of Syx17 (D), usnp (E), or VAMP7 (F) in LAMP1-GFP–marked cells blocks starvation-induced punctate LysoTracker red (LTR) staining. (G–J) Starvation leads to formation of LTR dots in control larvae (G). No LTR punctae form in starved Syx17 mutants (H), whereas LTR staining is restored in mutants expressing a Syx17 transgene (I). No LTR dots appear in VAMP7 mutants (J). (K) Expression of a second, independent usnp RNAi transgene also blocks LTR puncta formation. (L–O) Tandem-tagged mCherry-GFP-Atg8a is transported to autolysosomes that appear as mCherry-positive puncta (magenta in L) in control cells of starved larvae. Silencing of Syx17 (M), usnp (N), or VAMP7 (O) results in the formation of numerous dots positive for both mCherry and GFP (white). Dot plots in L′–O′ show intensity and colocalization profiles of mCherry and GFP dots. Pearson correlation coefficients shown at the top indicate strong colocalization of GFP and mCherry in M′–O′. (P–R) Quantification of data presented in A–C (P), D–F and K (Q), and G–J (R); n = 10 for all genotypes. mCh, mCherry. Error bars mark SDs. *, P < 0.05; ***, P < 0.001. Bar, 20 µm.