Fig. 6

YAP participates in thrombin-stimulated IL-8/CXCL8 expression in A549 cells. A A549 cells were transfected with YAP siRNA or control (con siRNA) for 48 h before being treated for additional 24 h with thrombin (10 U/ml). ELISA was performed to measure the level of IL-8/CXCL8 release (n = 3, mean ± SEM, *p < 0.05 vs thrombin-stimulated group). B A549 cells were transfected with control (con siRNA) or YAP siRNA for 48 h (n = 3, mean ± SEM, *p < 0.05 vs control). Immunoblotting was performed to determine the expression level of YAP. C. A549 cells were stimulated by thrombin (10 U/ml) for 0–60 min. Immunoblotting was performed to determine the expression level of phospho-YAP Ser127 (n = 4, mean ± SEM, *p < 0.05 vs non-stimulated cells). D Representative immunofluorescence images showing YAP (red) nuclear localization after 30 min thrombin (10 U/ml) stimulation in A549 cells. Cell nuclei were stained with DAPI (blue) (n = 3, original magnification = ×20). E Quantitative analysis of the percent of cells with YAP nuclei positive staining without (control) or with thrombin stimulation in A594 cells (n = 3, mean ± SEM, *p < 0.05 vs control). F A549 cells were transfected with control (con siRNA) or DCLK1 siRNA for 48 h before being stimulated for another 24 h with thrombin (10 U/ml). Immunoblotting was performed to determine the expression level of phospho-YAP Ser127 (n = 3, mean ± SEM, *p < 0.05 vs thrombin-stimulated cells). G A549 cells were incubated with Rhosin 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)