Fig. 2: Crizotinib inhibits TGFβ signaling in an ALK/MET/RON/ROS1-independent manner.

a–d NCI-H3122 cells were transfected with the indicated siRNAs for 48 h and further treated with or without TGFβ (1 ng/mL) for 30 min prior to western blot analysis. e, f A549 cells were treated with TGFβ (1 ng/mL), crizotinib (the indicated concentrations or 10 μM), or both for 30 min or the indicated times prior to western blot analysis. g A549 cells were treated with TGFβ (5 ng/mL), crizotinib (1 or 10 μM), or both for 24 h prior to western blot analysis. h Calu-1 cells were transfected with a pGL2-3TP-luciferase plasmid for 24 h and further incubated with TGFβ (1 ng/mL), crizotinib (the indicated concentrations), or both for 24 h. Luciferase activity was expressed as a relative value compared to that of the untreated cells, which was set to 100%. The data are expressed as the mean ± SEM (n = 6). **p < 0.01 and ***p < 0.005. i Calu-1 cells were treated with TGFβ (1 ng/mL), crizotinib (the indicated concentrations), or both for 30 min prior to western blot analysis. j Calu-1 cells were treated with TGFβ (5 ng/mL), crizotinib (1 or 10 μM), or both for 24 h prior to western blot analysis. k–n A549 cells were treated with TGFβ (1 ng/mL), ALK-specific inhibitors (1 or 10 M), MET-specific inhibitors (1 or 10 μM), or two of them for 30 min prior to western blot analysis.