NF-κB is dispensable for the loss of barrier function in LD-TNF/DN-Csk cells.

<p>Cells were seeded at confluence and incubated for 72 hours. Then, growth media was replaced by low serum media and cells were infected with adenovirus to express either LacZ (control) or DN-Csk. After another 16 h, cells were pretreated with DMSO (vehicle) or BAY 11–7082 (NF-kB inhibitor) for 30 minutes prior to TNF-α addition. <b><i>A</i></b>, Quantification of three independent Western blot experiments demonstrating that BAY 11–7082 is able to prevent ICAM-1 expression in LD-TNF/DN-Csk cells after 8 h of treatment with TNF-α. <b><i>B</i></b>, TEER measurements on ECIS electrodes. Notice the lack of effect of BAY 11–7082 treatment on the loss of TEER in LD-TNF/DN-Csk cells. Results are representative of three independent experiments performed in duplicate. Data presented as mean±SEM. Bar graph (mean±SEM) and statistical analysis of TEER for the 20 h time point corresponding to B is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161975#pone.0161975.s006" target="_blank">S6F Fig</a>.