DFO and Fer-1 respectively inhibited homocysteine-induced ferroptosis in cardiomyocytes.

(A) Western blots showed protein levels of β-catenin, active β-catenin, GPX4 and FTH1 in cardiomyocytes of each group as indicated. (B-E) Quantitative data on the abundance of specific proteins of Fig 3A were presented in indicated groups. *** P < 0.001, ** P < 0.01, * P < 0.05 vs the controls (n = 4). (F-H) Quantitative determination of Fe 2+ , lipid ROS 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). (I) Representative micrographs showed staining for β-catenin and GPX4 proteins in cardiomyocytes. Upper panel, immunofluorescent staining for β-catenin in given groups; Bottom panel, immunostaining for GPX4 in cardiomyocytes as indicated groups. Scale bar, 25 μm.