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. Author manuscript; available in PMC: 2013 Jul 25.
Published in final edited form as: Cytokine. 2012 Nov 21;61(2):608–613. doi: 10.1016/j.cyto.2012.10.018

Fig. 3.

Fig. 3

Functional analysis of putative NF-κB binding sites in HoxC4 promoter activity. (A) Effect of APRIL on the promoter activity of mutant HoxC4 promoter reporters. Three different mutant reporters were constructed (mt1, mt2, mt3), which lacked each NBE, as indicated. CH12F3-2A cells were transfected with the reporters (15 μg). APRIL was added, and the luciferase activity was determined 16 h later. Transfection efficiency was normalized to β-gal activity. Data shown are the average luciferase activity of three independent transfections with SEM (bars). *p < 0.05. (B) ChIP assay to determine whether NF-κB bound to the HoxC4 promoter region (NBE2). PCR of crosslinked chromatin precipitated from APRIL-stimulated or -nonstimulated (nil) CH12F3-2A and mouse spleen B cells with preimmune control (irrelevant rabbit IgG) or anti-p65 or anti-p50 Ab that were amplified for the HoxC4 promoter with the indicated primers.