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. 2024 Sep 23;14(16):6161–6184. doi: 10.7150/thno.98172

Figure 6.

Figure 6

ThA inhibits ferroptosis by activating the AMPK/Nrf2/GPX4 pathway. (A) Representative Western blot images of p-AMPK, AMPK, and GAPDH in PC-12 cells treated with ThA at specified concentrations. Full-length Western blot images are presented in Figure S34. (B) Bar chart indicates p-AMPK to AMPK ratio. (C) Representative Western blot images of p-AMPK, AMPK, nuclear Nrf2, GPX4, Lamin-B1, and GAPDH in PC-12 cells treated with ThA in the presence or absence of 5 μM CC. Full-length Western blot images are presented in Figure S35. (D-F) Bar charts indicate the ratios of nuclear Nrf2 to Lamin-B1, p-AMPK to AMPK, and GPX4 to GAPDH. (G) Representative immunofluorescence images demonstrate Nrf2 nuclear translocation and GPX4 expression in PC-12 cells treated with 8 μM ThA in the presence or absence of 5 μM CC. Magnification: 63×, Scale bars: 50 μm. (H) Bar chart indicates the mean fluorescence intensity of Nrf2 nuclear translocation and GPX4 expression in PC-12 cells. (I) Bar chart indicates viability of PC-12 cells cotreated with RSL-3 and CC in the presence or absence of ThA and Lip-1 at specified concentrations. (J) Representative images of DCFDA-stained PC-12 cells treated with 0.5 μM RSL-3 and 5 μM CC in the presence or absence of 8 μM ThA and 0.2 μM Lip-1. Magnification: 10×, Scale bars: 5 μm. (K) Bar chart indicates DCFDA intensity. (L) Representative images of C11 BODIPY-stained PC-12 cells cotreated with 0.5 μM RSL-3 and 5 μM CC in the presence or absence of 8 μM ThA and 0.2 μM Lip-1. Magnification: 10×, Scale bars: 200 μm. (M) Bar chart indicates the C11 GFP/RFP ratio. Bar, SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001.