Fig. 3

TSPAN9 increases the level of autophagy in gastric cancer cells. a AGS, AGS/R, MGC803 and MGC803/R cells were treated with 100 µM 5-FU for 48 h and then subjected to TEM. Autophagosomes are indicated by red arrows. b AGS, AGS/R, MGC803 and MGC803/R cells were treated with 100 µM 5-FU for 48 h followed by immunoblotting with the indicated antibodies. c AGS, AGS/R, MGC803 and MGC803/R cells transduced with GFP-LC3 plasmid were treated with 100 µM 5-FU for 24 h. After the cells were stained with DAPI, the fluorescence of the cells was detected under a confocal microscope. d Western blotting was used to assess the levels of autophagy-associated proteins after the transfection of TSPAN9 in AGS cells and TSPAN9-shRNA in AGS/R cells. e Western blotting was used to assess the levels of autophagy-associated proteins after the transfection of TSPAN9 in MGC803 cells and TSPAN9-shRNA in MGC803/R cells. f After transfecting TSPAN9 into AGS cells and TSPAN9-shRNA in AGS/R cells infected with GFP-LC3 plasmid, the cells were treated with 100 µM 5-FU for 24 h. The cells were then stained with DAPI, and the fluorescence was detected under a confocal microscope