Fig. 3

Involvement of c-Src in TNF-α-induced VCAM-1 expression. a, b Cells were pretreated with (a) various concentrations of PP1 (c-Src inhibitor; 0.1, 0.3, or 1 μM) (B) 1 μM PP1 for 1 h and then incubated with TNF-α for 16 h (a) or 4 h (b, c) Time dependence of TNF-α-induced c-Src phosphorylation, cells were treated with TNF-α (15 ng/ml) for the indicated time intervals in the presence or absence of c-Src siRNA. d Cells were transfected with siRNA of c-Src or scramble (scrb, as a control) for 24 h and then incubated with TNF-α for 16 h. e Cells were treated with TNF-α for the indicated time intervals in the presence or absence of TNFR nAb (10 μg/ml). The VCAM-1 protein (a, d), and mRNA and promoter activity (b) were analyzed by Western blotting, real time-qPCR, and promoter assay, respectively. The c-Src phosphorylation (c, e) was analyzed by Western blot. Data are expressed as mean ± SEM of three individual experiments (n = 3). ^* P < 0.05, ^# P < 0.01 vs. TNF-α alone