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For immunostaining, cells were permeabilized for 1 min using ice-cold PBS supplemented with 0.1% Triton X-100, blocked for 30 min with PBS supplemented with 1% BSA, incubated with the primary antibody in blocking buffer for 2 h at room temperature (RT), washed and subsequently incubated with secondary antibodies and DAPI in blocking buffer for 2 h at RT

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The first starves cells of cystine by inhibiting system Xc , collapsing the GSHGPX4 axis and predisposing lesions to lipid peroxidation

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