The miR-130a target site in the FOG-2 3′UTR is required for translational repression.

<p>In (A), a schematic of the constructs used to evaluate the function of the conserved region of the FOG-2 3′ UTR. In (B), NIH 3T3 fibroblasts were transfected with the constructs shown above along with pVRβgal. Forty-eight hours post transfection, cell lysates were assayed for luciferase activity and normalized to β-galactosidase activity. Results reported are the mean±S.E.M. (n = 8). In (C), northern analysis of 10 µg total RNA from transfected fibroblasts from (B) using a probe specific to the luciferase coding region (above) or β-galactosidase (below). In (D), primary neonatal cardiomyocytes were transfected with pVRβgal and a luciferase reporter containing the 3′UTR of FOG-2 or the ΔA 3′UTR mutation. Forty-eight hours after transfection, cells were assayed for luciferase and β-galactosidase activity. Results are reported as the mean normalized luciferase activity±S.E.M. (n = 20). In (E), NIH 3T3 fibroblasts were transfected with a luciferase reporter containing the 3′ UTR of FOG-2 (columns 1–3) or the ΔA mutation (columns 4 & 5) in the absence (columns 1 & 4) or presence of increasing amounts of 2′-O-methyl oligonucleotide (columns 2, 3, 5). Forty-eight hours post transfection, cell lysates were assayed for luciferase activity and normalized to β-galactosidase activity. Results reported are the mean±S.E.M. (n = 7). ‘*’ indicates a statistically significant difference (p<0.01 )