FIGURE 6.

Sp1 binds the miR-141 and miR-146b-5p promoters and activates their transcription in a p16-dependent manner. A, ChIP assay. Chromatin was purified from HFSN1C and HFSN1p16sh, and then immunoprecipitated (IP) using anti-Sp1 antibody. miR-141 and miR-146b-5p promoters were amplified by PCR (upper panel) and qPCR (lower panel) using specific primers, and their abundances were plotted relative to the input, and GAPDH (unlinked locus) was used as negative control. These experiments were performed in triplicate (*, p < 0.05). B, ChIP assay. The promoters of the indicated genes were amplified using qPCR. C, total RNA was extracted from HFSN1 cells expressing either Sp1-shRNA (scrambled sequence was used as control) or Sp1-ORF (empty vector was used as control) and was utilized to assess the expression level of the indicated genes by qRT-PCR. Error bars represent means ± S.D.