Fig. 7

Thrombin induces DCLK1, RhoA, and YAP activation through the ERK pathway. A A549 cells were incubated with U0126 for 30 min prior to stimulation for another 20 min with thrombin (10 U/ml). Immunoblotting was performed to determine the expression level of phospho-DCLK1 Ser30 (n = 4, mean ± SEM, *p < 0.05 vs thrombin-stimulated cells). B A549 cells were incubated with U0126 (30 min) prior to thrombin (10 U/ml) treatment for another 5 min. A Western blot pulldown assay was performed to determine RhoA activity (RhoA-GTP) level. Total RhoA level was used as a loading control (n = 3, mean ± SEM, *p < 0.05 vs thrombin-stimulated cells). C A549 cells were incubated with U0126 for 30 min prior to thrombin (10 U/ml) treatment for another 5 min. Membrane fraction was immunoblotted to determine the expression level of RhoA (n = 3, mean ± SEM, *p < 0.05, vs thrombin-stimulated cells). D A549 cells were preincubated with U0126 for 30 min prior to treatment with thrombin for another 30 min. Immunoblotting was performed to determine the expression level of phospho-YAP Ser127 (n = 3, mean ± SEM, *p < 0.05, vs thrombin-stimulated cells)