Figure 1. AMF induces PI3K/MAPK signaling pathways in a HER family dependent manner in T47D cells.
A, Cells serum-starved for 16 hr were stimulated with purified AMF, p-AKT and p-ERK levels were monitored in dose-dependent treatment for 15 min and B, for 5 to 30 min with AMF (90 nM). C, Suppression of AMF-induced p-AKT/p-ERK by 100 nM Wortmannin (PI3K inhibitor) and 20 μM U0126 (MAPK inhibitor). Serum-starved cells were pretreated with inhibitors for 1hr before AMF stimulation for 15 min. D, Effect of Lapatinib, dual tyrosine kinase inhibitor (10 μM) on p-AKT/p-ERK expression. Serum starved cells pretreated with Lapatinib for 1hr and stimulated by AMF (90 nM) or EGF (16 nM) for 15 min. Control indicates bovine serum albumin. E, After pretreatment with broad spectrum MMPs inhibitor (50 μM, Marimastat) for 1hr or 0.2 ng/ml pertussis toxin (G protein transduction inhibitor) for 16 hr, the cells were stimulated with AMF or lysophosphatidic acid (LPA, 10 μM), a GPCR stimulator for 15 min.