FIGURE 5.

Individual roles for RelA and c-Rel on LPS-induced Igκ gene transcription and pair-wise enhancer interactions. A, Western blot of RelA, actin and c-Rel in 70Z/3 cells infected with Luci or RelAi shRNA-expressing lentiviruses. B, Western blot of c-Rel and actin in splenic B cells from wild type and c-Rel knockout mice. C, Real-time RT-PCR analysis of Igκ gene transcript levels before and after LPS treatment for 20 h in control or RelA knock-down 70Z/3 cells. Standard deviations of two independent RNA purifications are indicated. D, Real time RT-PCR analysis of Igκ gene transcript levels in 48 h LPS-stimulated splenic B cells from wild type and c-Rel knockout mice. Standard deviations of two independent RNA purifications are indicated. E, 3C assays of control and RelA knock-down 70Z/3 cells treated with or without LPS as above. F, 3C assays of splenic B cells from wild type or c-Rel knockout mice treated with or without LPS as above. Standard deviations of two independent chromatin preparations are indicated. G, Real time PCR ChIP assays of RelA in control or RelA knock-down 70Z/3 cells treated with or without LPS as above. Fold enrichment refers to the sequence abundance in the immunoprecipitated sample divided by the corresponding sequence abundance in input DNA. Standard deviations of two independent chromatin preparations are indicated. H, Real time PCR ChIP assays of c-Rel in control or RelA knock-down 70Z/3 cells treated with out without LPS as above. Fold enrichment is defined in Figure 5G. Standard deviations of two independent chromatin preparations are indicated. I, Real time PCR ChIP assays of RelA in wild type or c-Rel knockout splenic B cells treated with or without LPS as above. Fold enrichment is defined in Figure 5G. Standard deviations of two independent chromatin preparations are indicated.