Figure 2

Reduction in levels of trimethylation of histone H3 lysine 4 (H3K4me3) and mRNAs of cytokine and chemokine genes in RASFs upon small interfering RNA (siRNA)-mediated inhibition of MLL1. (A) MLL1 mRNA levels in RASFs treated with control siRNA or MLL1 siRNA, as determined by quantitative RT-PCR (n = 12 RA patients). (B) Representative MLL1 protein expression patterns in RASFs treated with control siRNA or MLL1 siRNA by western blotting analysis. Vinculin was used as the internal control. Original blots are presented in Supplementary Figs. 2A and B. (C) The quantified data of MLL1 protein levels are presented as mean ± SEM (n = 7 RASFs that were treated with control siRNA or MLL1 siRNA). The intensity of MLL1 was normalized based on Vinculin. (D) H3K4me3 levels of interleukin (IL)-6, IL-15, C–C motif chemokine ligand (CCL)2, CCL5, C-X-C motif chemokine ligand (CXCL)9, CXCL10, CXCL11, C-X3-C motif chemokine ligand (CX3CL)1, and CXCL12 genes in RASFs treated with control siRNA or MLL1 siRNA, as determined by quantitative chromatin immunoprecipitation (ChIP)-PCR (n = 8 RA patients). (E) IL-6, IL-15, CCL2, CCL5, CXCL9, CXCL10, CXCL11, CX3CL1, and CXCL12 mRNA levels in RASFs treated with control siRNA or MLL1 siRNA, as determined by quantitative RT-PCR (n = 12 RA patients). Bars show the mean ± SEM. Values are expressed as the fold-increase versus the value for control siRNA-treated RASFs. * = P < 0.05; ** = P < 0.01; *** = P < 0.001 by Wilcoxon signed-rank test. See Fig. 1 for other definitions.