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Semi-automated freeze-substitution was performed using an AFS2 device (Leica) equipped with customized aluminum sapphire disc revolvers and samples were subsequently embedded in epoxy resin according to a previously published protocol 73

Loss of BRCA1 function caused by BRCA1 mutation results in the production of hydrogen peroxide in both epithelial breast tumor cells and adjacent stromal fibroblasts, which is able to promote the onset of a reactive glycolytic stroma, suggesting the metabolic phenotype of stromal cells in the TME may also be affected by BRCA1 mutation in tumor cells (225)