Figure 4

The involvement of PAR-1 in laminar flow-induced expressions of KLF2- and Nrf2-dependent genes. HUVECs were transfected with 100 pmol/L of control siRNA or PAR-1 siRNA for 2 days and then exposed to laminar flow for 24 hours. (A) Protein levels were determined by immunoblotting with specific antibodies against eNOS, NQO1, HO-1, PAR-1, and anti-tubulin. (B–F) mRNA levels of eNOS, NQO1, ferritin heavy chain (Ferritin H.C) and PAR-1 was determined by RT-qPCR. Relative expression levels were normalized versus GAPDH. Results are representative of three independent experiments that yielded similar results. Results are presented as the means ± SDs (n = 3). ^*P < 0.05; ^**P < 0.01 compared with control siRNA transfected HUVECs. ^#P < 0.05; ^##P < 0.01 compared with control siRNA transfected HUVECs under laminar flow conditions (12 dynes/cm²). Cells were used for experiments at passage 4, 5 and 6.