Bay11-7082 inhibits pro-IL-1β production in response to P. aeruginosa 
In addition to inhibition of NF-κB activity, Bay11-7082 can also directly inhibit the NLRP3 inflammasome [38]. To validate its use in this study as an NF-κB inhibitor, western blots for pro-IL-1β were performed alongside inhibition of CFTR activity by CFTRinh172 in response to PAO1 at 4 hours after stimulation (Fig. 7a). Our results indicate that Bay11-7082 prevents production of pro-IL-1β whereas CFTRinh172 does not seem to affect it. This was further corroborated by the ability Bay11-7082 to inhibit IκBα degradation at 0.5, 1, and 1.5 hours post PAO1 stimulation (Fig. 7b).
10.1371/journal.pone.0037689.g007 Figure 7  Bay11-7082 inhibits pro-IL-1β production in response to P. aeruginosa.
PMA-differentiated THP-1 cells were treated with 10 µM CFTRinh172 or 20 µM Bay11-7082 and harvested after (A) 4 hours (n = 3) or (B) 0.5, 1, and 1.5 hours (n = 3) stimulation with PAO1. One representative blot is shown with a graph of the averaged fluorescence intensity values over 3 experiments.

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