Fig. 4

BDH2 induces autophagy in GC cells. a Expression levels of LC3-II and p62 in BDH2-overexpressing cells were detected by western blotting. b TEM images of endogenic autophagic microstructures in BDH2-overexpressing and vector control cells. Red arrows indicate autophagosomes or autolysosomes. Scale bars, 1 μm. c Formation of endogenous LC3 puncta was detected by fluorescence microscopy. Scale bars, 20 μm. d, e BDH2-overexpressing and vector control cells were infected with an mRFP-GFP-LC3 lentivirus. f, g Numbers of LC3 puncta (yellow puncta for autophagosomes, red puncta for autolysosomes) were calculated. h Expression of LC3B in tumours was detected by IHC. Scale bars, 50 μm. i Xenografted tumours were subjected to TEM. Results are presented as means ± S.D. (n = 3); *p < 0.05, **p < 0.01 compared with vector; ^##p < 0.01 compared with BDH2 transfected cells treated with Baf-A1