Fig. 4: BAP1 regulates ferroptosis through ACSL4.

A Protein levels of indicated genes in indicated 786-O cells were measured by Western blotting. n = 5. B Cell viability measured by CCK-8 assay in indicated cell lines treated with IKE (10 μM) for 24 h. C Lipid peroxidation was assessed by flow cytometry after C11-BODIPY staining in the indicated cells treated with IKE. D Cell death was measured in BAP1-KO 786-O cells restored with ACSL4 treated with IKE (10 μM) for 24 h. E Cell viability measured by CCK-8 assay in the indicated cell lines cultured in medium with different concentrations of RSL3. F Bar graph showing cell viability in indicated cells treated with RSL3 (1 μM) combined with 2 μM ferrostatin-1 (Ferr-1) or 100 μM deferoxamine (DFO). G Cell viability measured by CCK-8 assay in the indicated cell lines cultured in medium with different concentrations of cystine. H Bar graph showing cell viability in indicated cells treated with cystine limitation combined with 2 μM Ferr-1 or 100 μM DFO. I Western blotting analysis of ACSL4 expression in the corresponding cell lines. n = 4. J Bar graph showing cell viability in indicated cells treated with cystine limitation combined with 2 μM Ferr-1 or 100 μM DFO. K Bar graph showing lipid peroxidation in the indicated cells cultured in cystine-free medium. L Cell death was measured in the indicated UMRC6 cells cultured in cystine-free medium for 48 h. Bar graph showing cell viability in indicated cells treated with 10 μM TBH (ROS inducer) with or without rosiglitazone (10 μM) (M) or abemaciclib (1 μM) (N). Error bars are mean ± SD. All P values were calculated using a two-tailed unpaired Student’s t-test. n ≥ 3 independent repeats unless specified.