Figure 5
SMYD3 is partially responsible for H3K4 trimethylation at the foxp3 locus in inducible regulatory T (iTreg) cells. (a) Western blots of SMYD3, H3K4me3, and β-actin from lysates of control or SMYD3 small interfering RNA (siRNA)-transfected wild-type (WT) naïve CD4+ T cells and from WT and SMYD3−/−-derived naïve CD4+ T cells, 4 days under iTreg-differentiating conditions. (b) Schematic of the murine foxp3 locus highlighting the regions investigated. Chromatin immunoprecipitation (ChIP) assays were performed in the foxp3 promoter site, CNS1, -2, and -3 of the foxp3 locus using H3K4me3 antibody or immunoglobulin G (IgG) as control (c), and in the foxp3 promoter site and CNS1 using SMYD3 antibody or IgG as control (d). (e) ChIP assays were performed in the foxp3 locus using H3K4me3 antibody or IgG as control or skewed T cells. Data show mean±s.e.m. of four replicate wells and are representative of three independent experiments. *P<0.05, **P< 0.01, and ***P<0.001. CNS, conserved noncoding DNA sequence.
