FIGURE 5.

ROCK2 is the dominant mediator of LPA-driven ICAM-1 and VCAM-1 mRNA and protein expression in HUVECs. siRNA-mediated gene knockdowns were performed as described in the legend to Fig. 3, and cells were then incubated with LPA for an additional 4 h. Knockdown efficacy was evaluated by Western blotting (A). ICAM-1 (B) and VCAM-1 (C) mRNA levels were measured by quantitative real-time PCR using gene-specific primers. Values are means (n = 4) ± S.E. ***, p < 0.0001 compared with nonspecific control (NC) siRNA treatment. D, following siRNA treatment and exposure to LPA for 4 h, ICAM-1 and VCAM-1 proteins were visualized by Western blotting with gene-specific antibodies. GAPDH was used as a normalization control.