Fig. 5

SiRNA against Poldip2 blocks LPS-induced p65 nuclear translocation. Confluent RBMVECs were transfected with siRNA against Poldip2 (siPoldip2) or control (siCtl) and treated with LPS (1 μg/mL) or PBS. a Following transfection and PBS or LPS treatment for 1 h, RBMVECs were fixed and incubated with antibodies against p65 (green) and DAPI (blue). White represents areas of colocalization. Scale bars equal 20 μm. Representative images are shown. b The areas of overlap between DAPI and p65 was measured by ImageJ. The graph depicts the average fold change in colocalization ± SEM. Bars represent mean ± SEM of 4 independent experiments. Two-way ANOVA ****p < 0.0001 vs. siCtl + PBS, ^####p < 0.0001 vs. siCtl + LPS. c After transfection and 1 h of treatment, RBMVECs were fractionated and the nuclear fraction was blotted for p65 and Lamin B1. Representative blots are shown. d The graph depicts nuclear p65 normalized to Lamin B1. Bars represent mean ± SEM of three independent experiments. Two-way ANOVA, ***p < 0.001 vs. siCtl, ^##p < 0.01 vs. siCtl + LPS