Figure 4.

TMEM74-induced autophagy is associated with ATG5-ATG12 complex, independent with BECN1-PIK3C3 complex. HeLa cells were firstly treated with the indicated siRNAs (siATG5 (a), siATG7 (b), siATG16L1 (c), siATG3 (d), siPI3KC3 (e), siATG10 (f), siBECN1 (g), siULK1 (h)) for 24 h, then transfected with GFP-TMEM74 or GFP for 24 h,meanwhile treated with or without bafilomycin A1 (100 nM) for at least 12 h. The levels of LC3B-II were detected by western blotting. (i) HeLa cells were transfected with GFP-TMEM74 or GFP (control) for 24 h respectively, treated with or without LY294002 (10 μM) for at least 8 h. The levels of LC3B-II were detected by western blotting