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. 2014 Feb 6;5(2):e1048. doi: 10.1038/cddis.2014.2

Figure 6.

Figure 6

miRNA let-7e targets MMP9 in ASCs during differentiation. (a) Untreated ASCs transfected with 100 nM let-7e (+let-7e) downregulate the expression of MMP9 mRNA measured by qRT-PCR; n=4. Data were normalized to GAPDH. Results are expressed as mean±S.D. *P<0.05 versus +Control. (b) Gelatin zymography of supernatants 24 h after transfection with let-7e (100 nM) or control oligonucleotides. (c) miRNA let-7e targets the mouse MMP9 3′UTR in HEK293 cells, (d) miRNA let-7e targets the mouse MMP9 3′UTR in untreated ASCs and in ASCs undergoing differentiation with ATRA. Cells were transfected either with the control vector (Control), a wild-type construct (3′UTR-MMP9) with an intact let-7e-binding site or the mutated construct (3′UTR-mutMMP9), as indicated. Cells were co-transfected with either control miRNA (c), miRNA let-7e (+let-7e), control antimiR (CantimiR) or anti-let-7e (+antiLet-7), respectively. Relative luciferase activity was measured by determining firefly and Renilla luciferase. Data are expressed as % of luciferase activity; n=4. Data are means±S.D. *P<0.05 versus Control