Fig. 3

Effect of DBPR114 on receptor tyrosine kinase phosphorylation in human HCC cells. A and C Modulation of MET receptor tyrosine kinase and downstream effector proteins. HA22T/VGH and Hep3B cells were treated with DBPR114 for 2 h, and 25 ng/mL HGF (ligand for MET) was added 10 min before the end of drug treatment. B and D Modulation of AXL and MERTK receptor tyrosine kinases. HA22T/VGH and Hep3B cells were treated with DBPR114 in the presence of 200 ng/mL GAS6 (ligand for AXL and MERTK) for 30 min. At the end of drug treatment, cells were lysed, and the soluble protein was separated using electrophoresis on a sodium dodecyl sulfate and polyacrylamide gel and analyzed through Western blotting. Autoradiographs were scanned for densitometric analysis using Image J software. Quantitation of the protein band was determined through normalization with the internal control GAPDH. Each bar represents the average value of three experiments. ND: not detected.  Representative gel images are presented from three independent experiments. *p < 0.05 vs. DMSO-treated cells, ^&p < 0.01 vs. DMSO-treated cells, ^#p < 0.001 vs. DMSO-treated cells, measured using  nonparametric t test