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. 2015 Jan 23;17(1):124–133. doi: 10.1016/j.neo.2014.11.009

Figure 4.

Figure 4

Expression of EGFR is diminished during the metastatic progression of human BCs. (A) The human MCF10A progression series consisting of normal (MCF10A), transformed (T1k), malignant (Ca1h), and metastatic (Ca1a) cells were serum deprived and stimulated with EGF for 30 minutes and subsequently analyzed for EGFR phosphorylation (pEGFR) and total expression of EGFR and Mig6. Actin served as a loading control. (B) Human MDA-MB-231 BC cells were engrafted onto the mammary fat pad of female nu/nu mice and resulting metastases from the lymph node and lungs were harvested and subcultured. These two independent cell lines and the parental cells were stimulated with TGF-β1 for 48 hours and analyzed for expression of EGFR and Mig6. Actin served as a loading control. (C) The murine NME progression series was analyzed by immunoblot for the expression EGFR and Mig6 under nonstimulated conditions (−) and following a 48 hour treatment with TGF-β1 (5 ng/ml). Data in A-C are representative of at least 3 independent experiments yielding similar results.