Increased β-catenin expression inhibited homocysteine-induced ferroptosis and upregulated expression of GPX4 in cardiomyocytes.

(A) Western blots showed protein levels of β-catenin, active β-catenin, GPX4, FTH1, SLC7A11 and ACSL4 in cardiomyocytes of each group as indicated. (B-G) Quantitative data on the abundance of specific proteins of Fig 4A 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 homocysteine stimulation alone (n = 4). (H-J) Quantitative determination of Fe 2+ , MDA and GSH 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 homocysteine stimulation alone (n = 4). (K) The cell viability was assayed with CCK-8 kit in each group. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls; ### P < 0.001, ## P < 0.01, # P < 0.05 vs homocysteine stimulation alone (n = 4). (L-M) Representative micrographs showed immunofluorescent staining for lipid ROS and GPX4 proteins in cardiomyocytes. Scale bar, 25 μm. (N) Representative transmission electron microscopy showed mitochondrial damage in cardiomyocytes. The yellow arrows indicated injured mitochondria. Scale bar, 300 nm.