Fig. 5

ATG7 inhibition induces MHC-I expression via the ROS/ NF-κB signaling pathway. (A-B) Effect of ATG7 on the protein expression or phosphorylation of NK-κB signaling components in LoVo and HCT-116 cells with shATG7 and ATG7-IN-1 (10 µM) treatment. (C-D) The CRC cells were treated with ATG7 inhibition (shATG7 or ATG7-IN-1, 10 µM, for 24 h), and the expression of p65 protein in both the nucleus and cytoplasm was assessed using western blot analysis. (E) Immunofluorescent staining detected the nuclear translocation of p65 in LoVo cells treated with ATG7 inhibition (shATG7 or ATG7-IN-1, 10 µM) (red: p65 positive stain, blue: nuclei positive stain, the arrows indicate the location of p65). (F) Intracellular ROS (green fluorescence) as detected by DCFH-DA staining (green). CRC cells were treated with ATG7 inhibition for indicated times and fluorescence images were captured. (G) GSH/GSSH ratio was measured in MSI CRC cells with shATG7 and ATG7-IN-1 treatment. (H-I) The impact of ATG7 inhibition (shATG7 or ATG7-IN-1, 10 µM) or NAC treatment (10 mM) for 1 h on the levels of p65 protein in both the nucleus and cytoplasm of CRC cells was evaluated. (J) Effect of ATG7 inhibition (shATG7 or ATG7-IN-1, 10 µM) or NAC treatment (10 mM) on MHC-I expression in CRC cells was determined by western blot. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001