Supplementary Figure 2: Induction of autophagy in response to ROS.

(a) Western analyses of MCF-7 stably expressing GFP-LC3 cells treated with 0.4 mM H₂O₂ for the indicated time with markers for autophagy (p62 and LC3), mTORC1 signaling (pS6K (T389), S6K, pS6 (S235/236) and S6). (b) Quantitation of the ratio of LC3 II/Actin from Fig. S2a. (±s.e.m., n = 3 independent experiments). ^*p<0.05,^***p<0.001, NS, not significant. (c) Western analysis of GFP-LC3 MCF7 cells pre-incubated in the presence or absence 100 nM Bafilomycin A1 (BafA1) for 1hr before treatement with 0.4 mM H₂O₂ for 7hr using anti-p62 and LC3 antibodies. (d) Quantitation of the ratio of LC3 II/Actin and p62/Actin from Supplementary Fig. S2c. (±s.e.m., n = 3 independent experiments). ^*p<0.05,^**p<0.01, NS, not significant. (e) Western analysis of Atg5^+/+ MEFs and Atg5^−/− MEFs treated with 0.4 mM H₂O₂ for 24hr using anti-p62 and Atg5 antibodies. Uncropped images of western blots are shown in Supplementary Fig. S6. Source data of statistical analysis are shown in Supplementary Table S1.