Research Paper Volume 14, Issue 6 pp 2678—2694

Figure 5. piRNA-6426 promotes SOAT1 promoter methylation by recruiting DNMT3B. The cells were transfected with LV-piRNA-6426 or together with LV-DNMT3B or sh-DNMT3B for 48 h. (A) RT-qPCR assay was used to detect the expression level of SOAT1 mRNA. (B, C) Western blotting was used to detect the expression levels of SOAT1 and DNMT3B proteins. (D) ChIP-qPCR was used to analyze the enrichment of DNMT3B in the SOAT1 promoter region after piRNA-6426 overexpression or interference, the normal rat IgG (IgG) was used as a negative control, data represent mean values relative to input (% Input). (E) MSP assay was used to detect the methylation level of SOAT1 promoter (the groups are consistent with panels B, C). Values were expressed as mean ± SEM. ns P>0.05, *P<0.05, **P<0.01, n=6.