Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study

Figure 3

Leukocyte binding to TNF-α-stimulated human retinal endothelial monolayers is significantly reduced by NF-κB1 silencing, but binding to untreated monolayers is not impacted. Human retinal endothelial cell monolayers were transfected with NF-κB1-targeted (NF-κB1) siRNA or nontargeted (NT) control siRNA and treated with TNF-α or no cytokine for 24 hours. Endothelial monolayers were incubated with THP-1 leukocytes for 20 minutes (1 × 10⁵ cells/7.5 mm² monolayer); monolayers were photographed, and the number of leukocytes bound per mm² was counted. (a) Graph showing leukocytes bound to the surface of endothelial monolayers. Bars represent mean number, with error bars showing standard error of the mean. monolayers/condition. Data were analyzed by two-tailed Student’s -test. ns = not significant. (b) Graph showing relative NF-κB1 transcript expression in human retinal endothelial cell monolayers that were transfected in parallel. Reference gene was 18S rRNA. Bars represent mean relative expression, with error bars showing standard error of the mean. monolayers/condition. Data were analyzed by two-tailed Student’s -test.

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