Breast Cancer and Fibroblast Cell Lines

MDA-MB-231, MDA-MB-468, and MCF-10A cells were purchased from the American Type Culture Collection (Manassas, VA, USA), and the mouse embryonic fibroblasts (MEFs) were received from Dr. Steven Frisch (West Virginia University). With regard to human mammary luminal epithelial (HMLE) cells, only cell lysates received from Dr. Alexey Ivanov (West Virginia University) were used. The conditions for cell growth were described previously [21 (link), 25 (link)]. Antibodies used in the study were anti-EGFR (610017) and anti-SHP2 (610822) from BD Biosciences (San Jose, CA, USA); anti-CBL (sc-1651), anti-pan-extracellular signal-regulated kinase 2 (anti-pan-ERK2; sc81457), and anti-ubiquitin (sc271289) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-β-actin (A5441) from Sigma-Aldrich (St. Louis, MO, USA); and anti-phospho-ERK1/2 (9101S), anti-phospho-Akt (9271S), and anti-pan-Akt (11E7) from Cell Signaling Technology (Danvers, MA, USA).