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Cells were rinsed with PBS and permeabilised with methanol at 20C for 10 min, before rinsing with PBS and blocking (1% milk (SigmaAldrich) in PBS) and incubating with anti-pERK antibody (1:150 Cell Signalling Technology 4370) in blocking solution for 1 h at room temperature followed by 3x PBS rinses and incubating with Hoechst 33342 (1g/mL Molecular Probes) and anti-rabbit AlexaFluor 546 or 568 (1:1000 Molecular Probes) in blocking solution for 1 h at room temperature

This suggests that ferroptosis, in conjunction with pyroptosis ( via NLRP3), may regulate atherosclerosis progression, offering insights into potential therapeutic targets for CAD (6)