Fig. 8. Additive effect of BITC and bafilomycin A1 on LC3B accumulation in AGS cells. (A) AGS cells were treated with BITC with or without 100 nM bafilomycin A1 for 24 h. Whole cell lysate was prepared, and the LC3B II level was measured by western blotting. β-actin was used as a loading control. (B) Increased LC3B protein expression was further confirmed by immunostaining with anti-LC3B antibody and was detected via Alexa Flour 488-conjugated secondary antibody (green) under a fluorescence microscope (scale-bar=50 μm). DAPI (blue) was used for nuclear staining. (C) The cytotoxicity of bafilomycin A1 was investigated by MTT assay after treating the cells with different concentrations of Bafilomycin A1 for 24 and 48 h. Data were calculated from three independent experiments and expressed as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 versus the control, and #p<0.05 versus BITC treatment alone.
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