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. 2018 Mar 30;22(6):3246–3258. doi: 10.1111/jcmm.13606

Figure 5.

Figure 5

HP1α inhibits DNMT3b activity (A) Co‐IP with anti‐DNMT3b followed by Western blotting with anti‐HP1α in hLCSCs transfected with pcDNA3.1‐DNMT3b. IgG IP served as negative control. Western blotting with DNMT3b served as INPUT. (B) ChIP assay with anti‐DNMT3b followed by PCR with telomere DNA primers in hLCSCs transfected with pcDNA3.1‐DNMT3b and pcDNA3.1‐HP1α. IgG ChIP served as negative control. PCR for telomere DNA served as INPUT. (C) TERRA promoter methylation analysis by MspI plus BamHI digestion in hLCSCs transfected with pcDNA3.1‐DNMT3b and/or pcDNA3.1‐HP1α. (D) TERRA promoter methylation analysis by Methylated DNA Immunoprecipitation (MeDIP)‐Dot blot‐Western blotting with anti‐5‐methylcytosine (5‐mC) in hLCSCs transfected with pcDNA3.1‐DNMT3b or/and pcDNA3.1‐HP1α. Each value was presented as mean ± standard error of the mean (SEM). mean ± SEM. **P < .01; *P < .05. For all Western blotting, we repeated the experiments for three times. We measured grey value of the bands for quantification. Each value was presented as mean ± standard error of the mean (SEM) (Student's t test)