Fig. 9

Effects of Set1 down-regulation on CD4⁺ T cells from 3 SLE patients. a, b Relative Set1 and CREMα protein expressions in SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA were quantified by western blot analysis 72 h after transfection. β-actin was used as endogenous control. c, d Relative Set1 (c) and H3K4me3 (d) enrichments at the CREMα promoter in SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA were analyzed by ChIP combined with qPCR 72 h after transfection. Input DNA was used as endogenous control, and IgG was used as negative control. e, f, g, h Relative levels of DNA methylation (e), DNMT3a (f), H3K9me3 (g), and SUV39H1 (h) at the CREMα promoter in SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA were evaluated by ChIP or MeDIP combined with qPCR 72 h after transfection. Input DNA was used as endogenous control, and IgG was used as negative control. i, j Relative IL-2 (i) and IL-17A (j) productions in the supernatants of SLE CD4⁺ T cells transfected with Set1-siRNA or control-siRNA were detected by ELISA 72 h after transfection. All experiments were repeated three times