Figure 1. Zyxin is required for WPB exocytosis mediated via cAMP pathway.

(a) ELISA of VWF secreted from HUVECs expressing scrambled shRNA (SCR) or shRNAs targeting 37 FA genes, with forskolin stimulation. The bar graph shows the ratio of secreted VWF to cytoplasmic VWF (n=12, *P<0.05, **P<0.01, ***P<0.001, versus scrambled shRNA, 3 independent experiments). (b) ELISA of VWF secreted from HUVECs expressing scrambled (SCR) or zyxin shRNAs (shZyxin-1 and -2) with stimulation of forskolin (left panel) (n=15, ***P<0.001, 3 independent experiments). The efficacy of zyxin knockdown was confirmed by western blotting analysis using an anti-zyxin antibody with ERK2 as a loading control (right panel). (c,d) ELISA of VWF secreted from HUVECs expressing scrambled (SCR) or zyxin shRNAs (shZyxin-1 and -2) with epinephrine (c) or thrombin (1 U ml⁻¹) and histamine (10 μmol l⁻¹) stimulation at the indicated concentration (d). (n=4, ***P<0.001). All error bars represent s.d. Student's t-test was used for statistical analysis. The uncropped scans of western blottings were shown in Supplementary Fig. 9.