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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 7. Misoprostol significantly increases CRE transcriptional activity and CREB binding to IL-10 promoter.

Figure 7

Cells were transfected with a CRE luciferase reporter vector as described in 2.9. The transfected cells were treated with Misoprostol (10 µM), H89 (10 µM) and LPS (100 ng/ml). Cytoplasmic extracts were prepared in 6 hours after LPS treatment and equal amounts were assayed for luciferase activity. ***P<0.001 compared to cells without Misoprostol treatment. Data are presented as means±SD, n=3. (B) RAW cells treated with and without Misoprostol were stimulated with LPS for indicated times. ChIP was performed with CREB antibody and IL-10 promoter was examined by qChIP PCR at CREB binding region (region I). *P<0.05 compared to UT without Mis, aP<0.01 compared to LPS, 30’ alone. (C) RNA Polymerase II ChIP was performed on cells treated as in (B), Il-10 promoter was examined by qChIP PCR at CREB binding region (I). **P<0.01 compared to UT without Mis, aP<0.05 compared to LPS, 30’ alone. (C) RNA Polymerase II binding to TSS site on IL-10 promoter was examined in cells treated as in (B) and (C) (II). *P<0.05 compared to LPS, 30’ alone. Data are presented as means±SD, n=3.