A subset of RelA:CBP/p300 dependent genes is independent of RelA(Ser276) phosphorylation.

<p>(A) Total RNA was isolated and purified from unstimulated cells at 30, 60, and 120 min of TNFα stimulation and prepared for analysis by RT-PCR. GAPDH was used as a reference and the 0 min time point for each cell line was used as the calibrator. The respective protein products of the genes (with names other than the gene name) are in parentheses. The <i>rela</i>⁻/⁻ cell line reconstituted with empty vector was included to ensure that RelA regulates the genes under study. The genes with their respective gene accession numbers (mouse) are as follows: <i>csf2</i> (NM_009969), <i>cxcl2</i> (NM_009140), <i>icam1</i> (NM_010493), <i>nfkbia</i> (NM_010907), <i>ptgs2</i> (NM_011198), <i>tnf</i> (NM_013693), <i>tnfaip3 variant 1</i> (NM_009397), <i>tnfaip3 variant 2</i> (NM_001166402), <i>tnfsf9</i> (NM_009404), and <i>vcam1</i> (NM_011693). Note that <i>tnfaip3</i> has two variants. (B) The RelA–TA2:TAZ1 interaction activates a set of RelA target genes independent of p-Ser276RelA. The expression phenotype was determined for 99 genes that undergo at least 2-fold activation at 1 h of TNFα stimulation as compared to unstimulated RelA(wt) reconstituted <i>rela</i>⁻/⁻ cells. These TNFα-activated genes were grouped into four groups, A–D, based on the observed expression defect (with 95% confidence) with respect to RelA(wt), in the two distinct RelA:CBP/p300 interaction defective mutant cells (see main text). The gene list was sorted according to the differences between the RelA(wt) and RelA(TA2) mutant reconstituted cells at 1 h of TNFα treatment. The genes that switch to other groups upon relaxation of the confidence limit to 67% are labeled with their prospective group names. The genes that were tested by qPCR are marked with a blue asterisk. <i>vcam1</i> did not satisfy the stringent criteria of at least 2-fold activation at 1 h of TNFα treatment and hence could not enter the list.