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Fig. 3. Vitamin C worked in a Jmjd2 dependent manner. (A) ChIP-qPCR of H3K9me3 at the IL17 promoter and enhancer (CNS2) locus. FACS-sorted naïve CD4+ T cells were cultured under Th17 conditions in the presence or absence of vitamin C and used for the ChIP analysis. Subsequent qPCR was done on DNA precipitated by control and anti-H3K9me3 Ab (mean ± SEM of duplicates, from one experiment representative of three independent experiments). (B) Quantitative RT-PCR was performed to assess the expression levels of Jmjd2 members in the indicated CD4+ T cells. (C) FACS-sorted naïve CD4+ T cells were transfected with control or Jmjd2a/b/c siRNAs, and then, the expressions of the indicated gene transcripts were checked by RT-qPCR. (D) The expression levels of Il17 (left) and Rorc (right) transcripts were investigated in the indicated Th17 cell populations. (E) IL17 expressions in the indicated Th17 cell populations were checked by flow cytometry. (F) ChIP-qPCR of H3K9me3 at the IL17 promoter and enhancer (CNS2) locus. FACS-sorted naïve CD4+ T cells were cultured under Th17 conditions in the presence or absence of vitamin C and used for the ChIP analysis. Subsequent qPCR was done on DNA precipitated by control and anti-H3K9me3 Ab (mean ± SEM of duplicates, from one experiment representative of three independent experiments). Data are representative of 3 (A, C-E) or 2 (B, F) independent experiments. **P < 0.01; *P < 0.05; NS, not significant.
BMB Reports 2017;50:49~54 https://doi.org/10.5483/BMBRep.2017.50.1.193
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