Fig. 5.

PLC-ε regulates NF-κB activity by promoting degradation of Iκ-Bα, nuclear DNA biding, and phosphorylation of RelA/p65. HPAEC were transfected with si-Con or si-PLC-ε. After 36 h, cells were treated with thrombin (5 U/ml) for 1.5 h. A: total cell lysates were prepared and immunoblotted with an anti-Iκ-Bα antibody. RelA/p65 levels were used to monitor loading. B: nuclear extracts were prepared and analyzed for RelA/p65 nuclear DNA binding using Cayman's NF-κB transcription factor assay kit as described in materials and methods. C: total cell lysates were immunoblotted with an anti-phospho-RelA/p65 (Ser⁵³⁶) antibody. RelA/p65 levels were used to monitor loading. Data are means ± SE (n = 6–8 for each condition). *P < 0.05 vs. si-Con untreated controls; #P < 0.05 vs. si-Con thrombin-treated controls.