Figure 2. Ferroptosis regulates cardiomyocyte cell death and LV remodeling following myocardial infarction.

(A) Measurement of total CK activity in control hearts and hearts subjected to 30 minutes of ischemia followed by 30 minutes of reperfusion using Langendorff preparations. Mice were treated with either vehicle control or Fer-1 two hours before harvesting. n = 5 per group. (B) Serial measurement of CK activity following reperfusion in control hearts and vehicle- and Fer-1–treated hearts subjected to IRI. n = 5 per group. *P < 0.05 versus control; **P < 0.05 versus other groups, by 2-sided Mann-Whitney U test. (C) Arachidonic acid metabolites measured by LC-MS/MS in control hearts and vehicle- and Fer-1–treated hearts subjected to IRI. Data are displayed as box-and-whisker plots. Lines indicate the mean value. *P < 0.05 versus control; **P < 0.05 versus other groups; ^#P < 0.05 versus the vehicle IRI group. (D) TTC staining and measurement of infarct area in hearts obtained from vehicle- and Fer-1–treated mice 48 hours following 90 minutes of IRI. *P < 0.05 compared with vehicle control. Original magnification, ×10. (E) Ly6G immunostaining and quantification 48 hours following 90 minutes of IRI in vehicle- and Fer-1–treated hearts. Blue indicates DAPI staining. Original magnification, ×200. *P < 0.05 versus vehicle control. (F) Echocardiography of vehicle- and Fer-1–treated mice 4 weeks following 90 minutes of IRI. Yellow dashed line indicates the akinetic area. (G–I) Quantification of LV ejection fraction, LV diastolic and systolic volumes, and akinetic area in vehicle- and Fer-1–treated mice 4 weeks after IRI. The akinetic area is expressed as a percentage of the LV area. *P < 0.05 versus vehicle control, by Mann-Whitney U test. (J) Picrosirius red staining (red) and infarct size quantification 4 weeks following 90 minutes of IRI in vehicle- and Fer-1–treated hearts. Lines represent the mean. *P < 0.05, by 2-sided Mann-Whitney U test.