Figure 6 Decreased autophagy and increased expression of ferritinophagy-related proteins in IAR-20 cells co-cultured with BMMSCs. (A) Western blot of LC-3, FTH1, NCOA4 and P62 in IAR-20 cells of different groups. Expression level of LC-3-II in the RB group was significantly lower than that in the R group and ROS group (ROS group vs Mock group, P<0.001; R group vs Mock group, P<0.001; R group vs ROS group, P=0.005; RB group vs Mock group, P<0.001; RB group vs ROS group, P<0.001; RB group vs R group, P=0.001). Expression level of FTH1 in the RB group was significantly higher than that in the R group and ROS group (ROS group vs Mock group, P=0.040; R group vs Mock group, P=0.014; R group vs ROS group, P=0.003; RB group vs Mock group, P=0.003; RB group vs ROS group, P=0.002; RB group vs R group, P=0.005). Expression level of NCOA4 in the RB group was significantly lower than that in the R group and ROS group (ROS group vs Mock group, P<0.001; R group vs Mock group, P<0.001; R group vs ROS group, P=0.013; RB group vs Mock group, P<0.001; RB group vs ROS group, P<0.001; RB group vs R group, P=0.002). Expression level of P62 in the RB group was significantly lower than that in the R group and ROS group (ROS group vs Mock group, P<0.001; R group vs Mock group, P<0.001; R group vs ROS group, P=0.863; RB group vs Mock group, P=0.081; RB group vs ROS group, P<0.001; RB group vs R group, P<0.001). $ P<0.05 vs the Mock group, * P<0.05 vs the ROS group, # P<0.05 vs the R group. (B) Western blot of LC-3, FTH1, NCOA4 and P62 in liver tissues of different groups. Expression level of LC-3-II in the BP group was significantly lower than that in the SCS group (SCS group vs Normal group, P<0.001; P group vs Normal group, P=0.017; P group vs SCS group, P<0.001; BP group vs Normal group, P=0.124; BP group vs SCS group, P<0.001; BP group vs P group, P=0.172). Expression level of FTH1 in the BP group was significantly higher than that in the P group and SCS group (SCS group vs Normal group, P=0.602; P group vs Normal group, P<0.001; P group vs SCS group, P<0.001; BP group vs Normal group, P<0.001; BP group vs SCS group, P<0.001; BP group vs P group, P=0.001). Expression level of NCOA4 in the BP group was significantly higher than that in the SCS group (SCS group vs Normal group, P<0.001; P group vs Normal group, P<0.001; P group vs SCS group, P=0.054; BP group vs Normal group, P<0.001; BP group vs SCS group, P=0.004; BP group vs P group, P=0.055). Expression level of P62 in the BP group was significantly higher than that in the P group and SCS group (SCS group vs Normal group, P<0.001; P group vs Normal group, P<0.001; P group vs SCS group, P=0.005; BP group vs Normal group, P=0.062; BP group vs SCS group, P<0.001; BP group vs P group, P<0.001). $ P<0.05 vs the Normal group, * P<0.05 vs the SCS group, # P<0.05 vs the P group. SCS – static cold storage; P – NMP; BP – BMMSCs combined with NMP; NMP – normothermic machine perfusion; BMMSCs – bone marrow mesenchymal stem cells; ROS – reactive oxygen species; LC-3 – light Chain 3; FTH1 – ferritin heavy chain; NCOA4 – nuclear receptor co-activator 4; P62 – sequestosome 1/p62.