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. Author manuscript; available in PMC: 2012 Mar 15.
Published in final edited form as: Arch Biochem Biophys. 2010 Dec 19;507(2):241–247. doi: 10.1016/j.abb.2010.12.020

Figure 3.

Figure 3

EMSAs confirm the binding of NF-κB to the −299 κB site. (A) VDR(+/−) MEFs were induced by TNF-α for 8 hours before nuclear extracts were prepared. Nuclear extracts were incubated with canonical κB probe (κBc), or −299 κB probe, and competed with 25-fold (25x) or 100-fold (100x) cold probe of its own or with cold mutant −299 κB probe or κBc probe. C, no nuclear extract control, P, probe. (B) VDR(+/−) MEFs were pre-incubated with ethanol (−) or 1,25(OH)2D3 (+) overnight, followed by 8 hours of PBS (−) or TNF-α (+) treatment. Nuclear extracts were incubated with −299κB probe. (C) TNF-α-treated MEF nuclear extracts were incubated with −299κB probe in the presence of anti-p65 (αP65) or anti-p50 (αP50) antibodies.