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. 2012 Apr;40(4):726–733. doi: 10.1124/dmd.111.040329

Fig. 5.

Fig. 5.

In vitro validation of SNP predictions. A, predicted miRNA interaction with wild-type and variant (rs11574744) HNF4A 3′-UTR. B, transfection of HeLa cell line with 200 ng of control pIS-0 plasmid or pIS-HNF4A or pIS-HNF4A_SNP constructs with the Renilla luciferase plasmid as internal control. Dual luciferase assays were performed at 24 h. Data are expressed as the pIS-HNF4A and pIS-HNF4A_SNP luciferase activity corrected for Renilla luciferase and normalized to pIS-0 within each experiment (mean ± S.E.M.). C, HeLa cells were cotransfected with 4 μg of pIS-HNF4A or pIS-HNF4A_SNP luciferase constructs, along with Renilla reporter plasmid for normalization. The cells were also transfected with miRNA Mimics (hsa-miR-34a or hsa-miR449a; 30 nM). All data are expressed as the pIS-HNF4A or pIS-HNF4A_SNP luciferase activity corrected for Renilla luciferase and normalized to the negative control (cel-miR-67) within each experiment (mean ± S.E.M.; n = 3 independent experiments). The assays were done in triplicate on three different days. *, p < 0.05.