Fig. 6. mAtg9 and autophagy. (A) HEK293/GFP-LC3 cells starved for 2 hours were labelled with antibodies for both endogenous Rab7 and mAtg9. White arrowhead points to GFP-LC3, mAtg9, Rab7 colocalization, blue to GFP-LC3 and mAtg9, and yellow to GFP-LC3 and Rab7. Bar, 10 µm. (B) HEK293/GFP-LC3 were treated with the siRNA for 72 hours, starved for 2 hours with ES medium or not (culture in FM), and extracted. Conversion of GFP-LC3-I to GFP-LC3-II was assessed using a GFP antibody. (C) Long-lived protein degradation was assessed 72 hours after siRNA addition in triplicate, as described in Materials and Methods. ^*P<0.01.