Figure 6

Amuvatinib suppresses MET receptor tyrosine kinase activity. (A) Flow cytometry analysis of p-MET (Tyr1234/1235) 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) MET staining in U266 cells treated with the indicated concentrations of amuvatinib (Rx) from quadruplet experiments as in (A). (C) U266 cells were serum starved and 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 MET (Y1349) phosphorylation. (D) The 140 kDa (solid bars) and the 170 kDa (speckled bars) phospho (black bars) and total (gray bars) MET bands from triplicate experiment as in (C) were quantitated and normalized to GAPDH levels. The results are presented as percentages of the HGF-stimulated DMSO controls. Data are representative of three independent experiments and presented as Mean ± SEM, n = 3, *P < 0.05, **P < 0.01.