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. Author manuscript; available in PMC: 2016 Dec 1.
Published in final edited form as: Clin Immunol. 2015 Sep 25;161(2):291–299. doi: 10.1016/j.clim.2015.09.008

Figure 5. Misoprostol significantly attenuates LPS inducible NFκB activity and p65 recruitment to TNF promoter.

Figure 5

(A) RAW cells were transfected with a luciferase reporter construct containing the NFκB-responsive IκB promoter. The transfected cells were pretreated with Misoprostol (10 µM) and stimulated with LPS (100 ng/ml) for 6 hours. Cytoplasmic extracts were prepared and equal amounts were assayed for luciferase activity. ***P<0.01 and aP<0.05 compared to LPS. (B) RAW cells treated with and without Misoprostol were stimulated with LPS for indicated times. ChIP was performed with p65 antibody and TNF promoter was examined by qChIP PCR at p65 binding region (region I). *P<0.05, **P<0.01 compared to UT without Mis, aP<0.05 compared to LPS, 15’ alone, bP<0.05 compared to LPS, 30’. (C) RNA Polymerase II ChIP was performed on cells treated as in (B), TNF promoter was examined by qChIP PCR at p65 binding region (regions I). *P<0.05, **P<0.01 compared to UT without Mis, aP<0.05 compared to LPS, 30’ alone. (D) RNA Polymerase II binding to TSS site on TNF promoter (region II) was examined in cells treated as in (B) and (C). *P<0.05 compared to UT without Mis, aP<0.05 compared to LPS, 30’ alone. Data are presented as means±SD, n=3.