Figure 4

Analysis of autophagy and mitochondrial dynamics in MEFs and hepatocytes in WT and Rev1^−/− cells. (a) Analysis of autophagy by immunoblot and (b) quantification of LC3B-II/Actin ratio in the liver of WT and Rev1^−/− 5 months old mice (females, n = 3; males n = 2). (c) Immunoblot analysis of total and phosphorylated DRP1 (Ser616) in the liver of WT and Rev1^−/− 5 months old mice (male and females together, n = 5). (d) Quantification of total and phosphorylated DRP1 (Ser616) from panel (c). (e) Immunoblot with WT and Rev1^−/− MEF cells, treated with rotenone (0 μM, 2 µM and 4 µM) for 12 hrs (n = 3; df = 2). (f) The LC3B-II/Actin ratio in untreated and treated MEF cells after 12 hrs (0 µM: t = 1, 2 µM: t = 2.2, 4 µM: t = 14). (g,h) Analysis of total DRP1 (0 µM: t = 0.56; 2 µM: t = 5.1; 4 µM: t = 7.36) and phosphorylated DRP1 (Ser616) (0 µM: t = 3.96; 2 µM: t = 21.93; 4 µM: t = 14.14) in untreated and treated MEFs. (i,j) Analysis of total AMPKα (0 µM: t = 2.09; 2 µM: t = 0.56; 4 µM: 81.52) and phospho- AMPKα (Thr172) (0 µM: t = 64.37; 2 µM: t = 21.2; 4 µM: t = 4.67) in untreated and treated MEFs. n = sample number; t = t value; df = degree of freedom; *p < 0.05. **p < 0.01. ***p < 0.001. ****p < 0.0001. Data presented are mean ± M.S.E. Dark bars: WT and gray bars: Rev1^−/−.