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. 2015 Apr 15;2015:746970. doi: 10.1155/2015/746970

Figure 4.

Figure 4

BRCA1 facilitates miR-143 and miR-145 processing. In order to confirm whether BRCA1 facilitates miR-143 and miR-145 processing, BRCA1 overexpression and siBRCA1 expression plasmids were transfected into MCF-7 cells. The empty vector or scramble siRNA expression vector served as controls. (a) BRCA1 mRNA level and (b) protein level were examined by qPCR and Western blot, respectively, in stable transfected cells, normalized by beta-actin (∗ P < 0.05 as compared with mock control; n = 3). Then, the expression levels of the primary (pri), precursor (pre), and mature (mat) forms of the indicated miRNAs were examined in human BRCA1 (c) overexpressed and (d) knock-down MCF-7 cells using qRT-PCR analysis. Pri- and pre-miRNAs were normalized by beta-actin, and mature miRNA was normalized by U6 snRNA (∗ P < 0.05 as compared with mock control; n = 3). Meanwhile, in vivo monitoring assay of pri-miRNA processing in (e) BRCA1 overexpression or (f) knock-down MCF-7 cells carrying miR-143 or miR-145 at the 3′ untranslated region of the luciferase gene. The intensities were normalized by Renilla luciferase and are shown as fold induction as compared with an empty pmirGLO vector (∗ P < 0.05; n = 3). Error bars represent standard deviation.