Fig. 4

Effects of Set1 down-regulation on CD4⁺ T cells from SLE patients. a, b Relative Set1 and CREMα protein levels were evaluated by western blotting analysis of SLE CD4⁺ T cells 72 h after transfection with Set1-siRNA or control-siRNA. β-actin served as an endogenous control. c, d Relative Set1 (c) and H3K4me3 (d) levels within the CREMα promoter in SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA were confirmed by ChIP and real-time PCR 72 h after transfection. Results were normalized to input DNA (total chromatin). e, f Relative IL-2 (e) and IL-17A (f) concentrations in the supernatants of SLE CD4⁺ T cells were measured by ELISA 72 h after transfection with Set1-siRNA or control-siRNA. g Relative DNA methylation level at the CREMα promoter in SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA was assayed by MeDIP and real-time PCR 72 h after transfection. h,i, j Relative enrichments of H3ac (h), H4ac (i), and DNMT3a (j) within the CREMα promoter region in SLE CD4⁺ T cells were tested by ChIP and real-time PCR 72 h after transfection with Set1-siRNA or control-siRNA. Results were normalized to input DNA (total chromatin). All experiments were performed in triplicate