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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Nature. 2010 Aug 26;466(7310):1115–1119. doi: 10.1038/nature09283

Figure 3. IκBβ is recruited to the promoter of TNFα together with P65 and c-Rel.

Figure 3

a,b, Raw264.7 were stimulated with LPS and immunoprecipitated (IP) with anti-IκBβ (a), anti-p65 (b) or anti-c-Rel (b) antibodies and immunoblotted (IB) as indicated. c, LPS-stimulated Raw264.7 lysates were immunoprecipitated with anti-IκBβ; eluted with IκBβ peptide; immunoprecipitated with anti-c-Rel antibody; and immunoblotted as indicated. d, Raw264.7 lysates were subjected to ChIP as indicated and analyzed by qPCR targeting TNFα and IL-6 promoter κB sites; error bars indicate ±s.d (n=3). e, ChIP was performed as in (d) on WT and IκBβ−/− BMDM treated with LPS for 2 hours; error bars indicate ±s.d (n=3). f, BMDM treated as in (e) were immunoprecipitated with anti-p65 antibody. g, RAW264.7 were treated with LPS and nuclear extracts were subjected to EMSA TNFα κB3 or κB2 probes. Super shifts were performed using cells stimulated for 1hr. h, BMDM were treated with LPS and EMSA and supershifts with the κB2 probe were performed as in (g).