Fig. 2

RSL3, a ferroptosis activator, accelerates ligation-induced neointima formation in mouse carotid arteries. A An experimental protocol. Two days after ligation, C57BL/6J mice were intraperitoneally injected with vehicle or RSL3 (10 mg/kg/day) for 14 days. Then, the carotid arteries were collected for further analysis. B Immunofluorescent staining of GPX4 in mouse carotid artery (neointima outlined by white dashed line). Quantification of GPX4 relative fluorescence intensity (n = 6 per group). Scale bar, 50 μm. C Perl’s Prussian blue staining in mouse carotid artery (neointima outlined by black dashed line). The number of iron⁺ cells in neointima per field (n = 6 per group). Scale bar, 100 μm. D Immunofluorescent staining of COX-2 in mouse carotid artery (neointima outlined by white dashed line). Quantification of COX-2 MFI (n = 6 per group). Scale bar, 50 μm. E H&E staining of ligated carotid arteries from C57BL/6J mice treated with RSL3 or vehicle (neointima outlined by black dashed line). The intima/media ratio was calculated (n = 6 per group). Scale bar, 100 μm. F Immunofluorescent staining of α-SMA in mouse carotid artery (neointima outlined by white dashed line). Quantification of α-SMA MFI (n = 6 per group). Scale bar, 50 μm. Data are presented as mean ± SEM, P values calculated by a two-tailed unpaired Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001