Fig. 6. GATE16 and GABARAP localize to the LC3-positive autophagosome under starvation conditions. (A) F9 cells stably transfected with CFP-LC3 (a,d), YFP-GATE16 (b,e), or YFP-GABARAP (c,f) were cultured under nutrient-rich conditions (a-c) or starvation conditions (d-f) for 2 hours at 37°C. Living cells were observed using a Delta Vision microscopic system. Scale bar, 10 µm. (B) F9 cells co-expressing YFP-GATE16 and CFP-LC3 (a,b), YFP-GABARAP and CFP-LC3 (d,e) were starved for 2 hours and subjected to fluorescence microscopic analysis. Merged images (c,f) are shown. Arrowheads indicate the co-localization of both the punctate and ring-shaped structures. Scale bar, 10 µm. (C) The C-terminal glycine is required for autophagosome-targeting of GATE16 and GABARAP. F9 cells expressing YFP-LC3^{C22, G120A} (a), YFP-GATE16^{C1, G116A} (b), or YFP-GABARAP^{C1, G116A} (c) were starved for 2 hours and subjected to fluorescence microscopic analysis. Scale bar, 10 µm.