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cDNA was synthesized with Superscript III (Invitrogen) and quantitative PCRs were done in triplicates with the GoTaq pPCR Master Mix (Promega) on a 7500 Fast Real-Time PCR System (Applied Biosystems)

Disrupting this network can simultaneously weaken parallel ferroptosis defense mechanisms, thereby lowering the threshold for lipid peroxidation and ferroptotic cell death in cancer cells

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However, in line with the specificity of CPA, the C-terminal residues of the L585I, L585M, and L585V variants were cleaved in vitro upon exposure to the enzyme for 5 hours at 37 C, as confirmed by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of the C-terminal peptides (Supplementary Table 1)