ThA inhibits ferroptosis by activating the Nrf2/Keap1 pathway. (A) Bar chartindicates PC-12 cell viability cotreated with RSL-3 and ML385 in the presence or absence of ThA and Lip-1 at specified concentrations. (B) Representative images of C11 BODIPY-stained PC-12 cells cotreated with 0.5 μM RSL-3 and 4 μM ML385 in the presence or absence of 8 μM ThA and 0.2 μM Lip-1. Magnification: 10×, Scale bars: 200 μm. (C) Bar chart indicates the C11 GFP/RFP ratio. (D) Representative images of DCFDA-stained PC-12 cells cotreated with 0.5 μM RSL-3 and 4 μM ML385 in the presence or absence of 8 μM ThA and 0.2 μM Lip-1. Magnification: 10×, Scale bars: 200 μm. (E) Bar chart indicates DCFDA intensity. (F) Representative images of PGSK-stained PC-12 cells treated with 100 μM hemin in the presence or absence of 8 μM ThA and 20 μM DFO. Magnification: 10×, Scale bars: 200 μm. (G) Bar chart indicates PGSK intensity in PC-12 cells. (H) Bar chart indicates viability of WT and Nrf2-/- HeLa cells treated with 0.5 μM RSL-3 in the presence or absence of 8 μM ThA. (I) Representative images of Hoechst/PI-stained WT and Nrf2-/- HeLa cells treated with 0.5 μM RSL-3 in the presence or absence of 8 μM ThA Magnification: 10×, Scale bars: 200 μm. (J) Bar chart indicates cell death of Hoechst/PI-stained HeLa cells. (K) Bar chart indicates viability of WT and Nrf2-/- HeLa cells treated with 100 μM hemin in the presence or absence of 8 μM ThA. (L) Representative Western blot images of Nrf2, GPX4, and GAPDH in WT and Nrf2-/- HeLa cells treated with or without 8 μM ThA. Full-length Western blot images are presented in Figure S33. (M, N) Bar charts indicate the ratios of Nrf2 and GPX4 to β-actin. Bar, SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001.