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4T1 breast cancer cells were previously derived from a spontaneously formed breast tumor in a BALB/c female mouse [21]. The 4TLM cell line was derived from liver metastases and 4TBM was derived from brain metastasis of heart metastatic subset of 4T1 cells by Erin et al. [2, 21, 22]. 4TLM and 4TBM cells were grown in DMEM-F12 medium supplemented with gentamicin and 5% FBS.
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