Effects of HRG on the phosphorylation of Akt and MAPK in tumor-derived cells

Copyright information:Taken from "Functional interaction between mouse erbB3 and wild-type rat c-in transgenic mouse mammary tumor cells"Breast Cancer Research 2005;7(5):R708-R718.Published online 6 Jul 2005PMCID:PMC1242139.Copyright © Kim et al. licensee BioMed Central Ltd. 85815 and 85819 cells were cultured overnight in medium containing 0.5% FBS before being exposed to HRG at the indicated concentrations for 2 h. Cells were harvested and 50 μg total cell lysates were subjected to Western blot analysis for total Akt, phosphorylated Akt, total ERK2 (polyclonal antibody C-14; Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA), and phosphorylated MAPK (E10 mAb; Cell Signaling Technology) expression. β-actin was used as loading control. 78423, 85815 and 85819 cells were cultured overnight in medium containing 0.5% FBS before being exposed to 2.5 ng/ml HRG for the indicated time intervals. At each time point, cells were harvested and 50 μg total cell lysates were subjected to Western blot analysis for total Akt, phosphorylated Akt, total ERK2, and phosphorylated MAPK expression. β-actin was used as loading control.