FIGURE 7. c-Rel binds TNFRSF13C genomic DNA endogenously and affects BAFF-R expression in B cell lines.

A, Western blot of the nuclear extracts of the plasma and B cell lines as well as normal primary peripheral blood (PB) B cells. Extracts were blotted for c-Rel and for Histone H1 as a nuclear control. B, Chromatin immunoprecipitation (ChIP) of the region upstream of the BAFF-R gene with c-Rel and Pax5 antibodies prepared from RAMOS cells. The enrichment values represent amounts relative to the amount of chromatin in the non-specific polyclonal Ab IP. C, Following c-Rel knockdown in Loukes and RAMOS cells with c-Rel-specific siRNA or GC-content-matched non-specific control siRNA, c-Rel and BAFF-R mRNA were measured with qRT-PCR. Amounts were calculated according to the ΔΔC_T method compared to 18S rRNA and normalized to expression in the cells treated with the GC-content control non-specific siRNA. Asterisks indicate a significant change in gene expression in the test c-Rel siRNA compared to control non-specific siRNA, p < 0.05, in three replicates.