Fig. 2

ERα repressed miR-29a expression at the transcriptional level. A Genome region of miR-29a at Chromosome 7q32.3. B ChIP-seq identified two ERE half sites in the promoter region and enhancer region of miR-29a. C Prediction of a conserved ERα binding motif -G/TGTCA- by CistromeDB Toolkit. D ChIP-PCR using MCF-7 cells to validate the two ERE half sites to the genome of miR-29a indicated in (B). IgG was used as a negative control, input was used for positive control. E Additional screening of ERα-binding sequences at the genome region of miR-29a by applying 12 pairs of primers to run PCR, followed by agarose gel electrophoresis (lanes 1–12). Primer pair 5 (P5) represented the predicted binding site at the enhancer region, and primer pair 7 (P7) represented the predicted binding site at the promoter region were indicated with red arrows. F, G Overexpression of ERα in MDA-MB-231 cells (F) suppressed the expression of miR-29a (G). H, I Knocked down of ERα in MCF-7 cells (H) promoted the expression of miR-29a (I). Data are presented as the mean ± SEM (n = 3). **p < 0.01