Fig. 3

TBM induces autophagy initiation in CRC cells. a. Immunoblot analysis of Atg5 expression in SW480 and HCT116 cells treated with indicated concentrations of TBM for 24 h. b. Immunoblot analysis of Beclin 1 expression in HCT116 and SW480 cells treated with indicated concentrations of TBM for 24 h. c. Immunoblot analysis of LC3 and Atg5 expression in CRC cells transfected with siNC or siAtg5 for 24 h, followed by treatment with or without 10 μM TBM for another 24 h. d. Immunoblot analysis of LC3 and Beclin 1 expression in HCT116 and SW480 cells transfected with siNC or siBeclin 1 for 24 h, followed by treatment with or without 10 μM TBM for another 24 h. e. SW480 and HCT116 cells were treated with vehicle, TBM (10 μM), 3-MA (5 mM), or in combination for 24 h. Immunoblot analysis was used to detect the expression of LC3. f. SW480 and HCT116 cells were transfected with GFP-LC3 for 48 h, and then were treated as in (E). The LC3 puncta by immunofluorescence analysis was shown and quantitated. Scale bar, 10 μm. g. Co-immunoprecipitation analysis of the interaction between Beclin1 and Bcl-2 in SW480 and HCT116 cells treated with or without 10 μM TBM for 24 h. ***, p < 0.001