Figure 7

Amuvatinib inhibits MET signaling. (A) Flow cytometry analysis of p-AKT (Ser473) levels in U266 cells were treated for 24 hrs with DMSO (gray shaded) and 25 μM amuvatinib (dark black line). (B) Quantification of phospho (black bars) and total (gray bars) AKT staining in U266 cells treated with the indicated concentrations of amuvatinib (Rx) from five experiments as in (A). (C) U266 cells were serum starved, treated with the indicated concentrations of amuvatinib or DMSO, and stimulated with 50 ng/ml HGF for 15 min. Cell lysates were subjected to immunoblot analysis to assess phosphorylation of ERK p44/42, AKT, and GSK-3β. AKT (D), GSK-3β (E), and ERK p44 (solid bars) and p42 (speckled bars) (F), phospho (black bars) and total (gray bars) bands were quantitated and normalized to GAPDH levels. The results are presented as a percentage of the HGF stimulated DMSO control. Data are representative of three independent experiments and presented as Mean ± SEM, n = 3, *P < 0.05.