GPX4 acted as a target gene of β-catenin.

(A) Western blots analysis showed protein levels of β-catenin and GPX4 in cardiomyocytes with overexpression or knockdown of β-catenin. (B-C) Quantitative determination of the abundance of specific proteins of Fig 5A were presented in indicated groups. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls (n = 4). (D) PCR analysis of GPX4 mRNA levels following β-catenin gene overexpression and silencing. (E) Quantitative determination of the abundance of GPX4 mRNA levels in Fig 5D were presented in indicated groups. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls (n = 4). (F) Dual luciferase reporter gene assay: both pGL6-TA and pRL-SV40-C were co-transfected into HEK293T cells with pcDNA3.1-β-catenin; both pGL6-TA and pRL-SV40-C were co-transfected into HEK293T cells in the control group. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls (n = 4). (G) ChIP assay verified that β-catenin can binded to the specific site on the promoter of GPX4 gene. (H) Western blots analysis showed protein levels of active β-catenin, β-catenin and GPX4 in cardiomyocytes of indicated groups. (I-K) Quantitative determination of the abundance of specific proteins of Fig 5H were presented in indicated groups. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls; ### P < 0.001, ## P < 0.01, # P < 0.05 vs β-catenin siRNA transfection alone (n = 4). (L-M) Quantitative determination of Fe 2+ and MDA in indicated groups on the colorimetric microplate reader. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls; ### P < 0.001, ## P < 0.01, # P < 0.05 vs β-catenin siRNA transfection alone (n = 4). Sc-siRNA, Scramble-siRNA.