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Expression of system x c was positively correlated with ER expression, and predicted the poor outcome of ER+ breast cancer To investigate how ER regulates ferroptosis in ER+ breast cancer cells, we conducted an analysis of ferroptosis-related genes that are also regulated by estrogen

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Five-millimeter sections were cut from the paraffin blocks and stained with cleaved caspase-3 (CST, Danvers, MA, USA, 9661, 1:200), PARP1 monoclonal antibody (Thermo Fisher Scientific Inc., Grand Island, NY, USA, 436400, 1:20), SQSTM1/p62 (CST, Danvers, MA, USA, 23214, 1:200), or LC3A/B (CST, Danvers, MA, USA, 12741, 1:400) at 4 C overnight

Subsequently, biotinylated detection antibodies specific to TNF- were added

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