Fig. 6
From: The IAP family member BRUCE regulates autophagosome–lysosome fusion
BRUCE interacts with an autophagosome-lysosome fusion regulator Syntaxin 17. a Co-immunoprecipitation of GFP-Syntaxin 17 (STX17) with Myc-BRUCE. GFP-STX17 and Myc-BRUCE WT were transfected in HEK293T cells. Myc-BRUCE was immunoprecipitated using anti-Myc antibody from total cell lysates. Myc-BRUCE and GFP-STX17 were detected by western blotting, using the indicated antibodies. *Nonspecific band. b Confocal microscopy images of Bruce+/+ and Bruce−/− MEFs, stably expressing exogenous GFP-STX17. Cells were grown in fully supplemented medium (FM) or starved for 2 h (Starv) with or without Bafilomycin (Baf; 100 nM). Scale bars, 20 µm. c Colocalization analysis of endogenous LC3B and exogenously expressed GFP- STX17. Bruce+/+ and Bruce−/− MEFs stably expressing GFP-STX17 were grown in FM or starved for 2 h with or without Bafilomycin (100 nM), fixed, and stained for endogenous LC3B. Line plots across 8 µm distance (marked with arrows) exemplify degree of colocalization of GFP-STX17 and LC3B signals. Scale bars, 20 µm
