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. Author manuscript; available in PMC: 2013 Apr 16.
Published in final edited form as: Cell Metab. 2011 Dec 7;14(6):747–757. doi: 10.1016/j.cmet.2011.11.006

Figure 2. TGR5 activation inhibits NF-κB activation via cAMP signaling.

Figure 2

(A) Western blot of C-Jun, phosphorylated c-Jun (P-C-Jun) with tubulin as loading control, and NF-κB p65 western blot of nuclear extract with PARP-1 as loading control of RAW264.7 macrophages treated with 100 ng/ml LPS for 3 hrs in combination with 100 μM SQ22536 and 30 μM INT-777 (n=3). (B) Quantification of western blot band intensity of p65 corrected for the intensity of PARP-1 using image analysis software. (C) Western blot of phosphorylated IκBα, total IκBα, and tubulin as loading control of lysate of RAW264.7 macrophages treated with 100 ng/ml LPS for 1 hr in combination with 30 μM INT-777 (n=3). (D) Quantification of western blot band intensity of IκBα corrected for the intensity of tubulin using image analysis software. (E) NF-κB-p65 binding activity to its DNA response element after 3 hours LPS stimulation. (F-H) LPS-induced (6 hrs) NF-κB transcriptional activity in RAW264.7 macrophages electroporated with the NF-κB reporter plasmid in combination with electroporation of TGR5 (F) or shTGR5 (G) in the presence of 30 μM INT-777 (black bars) or vehicle (white bars) (n=3) (H) LPS-induced NF-κB transcriptional activity in RAW264.7 macrophages electroporated with the NF-κB reporter plasmid in combination with 30 μM INT-777 treatment or vehicle in the presence of 100 μM SQ22536 (grey bars), 20 μM 2′, 5′-dideoxyadenosine (black bars) or control conditions (white bars) (n=3). Results represent the mean ± SEM. * Statistically significant, P<0.05.