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7AD), MDA-MB-231 cells from a highly aggressive, invasive and poorly differentiated triple-negative breast cancer (TNBC) cell line, were maintained in Dulbeccos modified Eagle media, Nutrient Mixture F12 (DMEM/F-12 media) with foetal bovine serum (5%), glutamine (2 mM), sodium pyruvate (1 mM) and non-essential amino acids (0.02 mM) at 37 C in a humidified atmosphere of 5% CO 2

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Brains were extracted and postfixed in 4% paraformaldehyde overnight, then transferred to 30% sucrose (Fisher Scientific) solution for cryoprotection and remained in the solution until sectioning