Fig. 7.

Hyperactive NRF2 inhibits RT-induced ferroptosis by regulating SLC7A11 in ESCC cells. a Western blotting and qRT-PCR analysis of NRF2 and SLC7A11 expression in NC and NF-OE KYSE 30 and KYSE 150 cells. b Western blot analysis of SLC7A11 levels in NF-OE KYSE 150 cells with SLC7A11 siRNA (NF-OE-SLC-Si), negative control (NF-OE-SLC-con), and NF-OE KYSE 150. c Lipid peroxidation assessment in NF-OE-SLC-Si and NF-OE-SLC-con KYSE 150 cells 24 h after exposure to 6 Gy of RT. Bar graph showing RT-induced relative fold changes of lipid peroxidation levels per 10,000 cells by C11-BODIPY staining in the indicated cells. d qRT-PCR analysis of PTGS2 expression in NF-OE-SLC-Si and NF-OE-SLC-con KYSE 150 cells 24 h after exposure to 6 Gy of RT. e Representative images of clonogenic survival assays in NF-OE-Con and NF-OE-SLC-Si KYSE 150 cells pre-treated with 5 μM ferrostatin-1 or DMSO for 24 h, followed by exposure to 4 Gy of RT, respectively. f The quantified clonogenic survival assay in NF-OE-Con and NF-OE-SLC-Si KYSE 150 cells that were pre-treated with 5 μM ferrostatin-1 or DMSO for 24 h followed by 4 Gy of RT, respectively. g Quantified cell proliferation assay curves analyzed by CCK-8 in NF-OE-SLC-con and NF-OE-SLC-Si KYSE 150 cells pre-treated with 5 μM ferrostatin-1 or DMSO for 24 h, followed by exposure to 6 Gy RT. Each experiment was conducted independently triplicate. Student’s t-test (two-tailed) was used for statistical analysis, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Bar graphs: mean ± SEM, n = 3