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. 2012 Aug 24;40(20):10478–10493. doi: 10.1093/nar/gks789

Figure 6.

Figure 6.

miR-101 regulates expression level of TGFB1 through AP-1. (A) The proposed miR-101-to-FOS-to-TGFB1 regulatory circuit. (B) Effect of miR-101 on FOS mRNA level. Cultured HepG2 were transfected with 15 nM of miR-101 or scrambled control for 48 h. FOS expression level was determined using qRT-PCR. ***P < 0.001. (C) Effect of miR-101 on TGFB1 expression level. Cultured HepG2 were transfected with 15 nM of miR-101 or scrambled control for 48 h. TGFB1 expression level was determined using qRT–PCR. ***P < 0.001. (D) Effect of miR-101 on FOS 3′-UTR and TGFB1 3′-UTR reporter activity. Cultured HepG2 were transfected with 15 nM of miR-101 or scrambled control for 24 h. Cells were cotransfected with plasmids containing firefly luciferase reporter carrying FOS 3′-UTR or TGFB1 3′-UTR and control Relina luciferase and incubated for additional 24 h. Luciferase activity was determined as described in materials and methods. The firefly reporter activity was normalized to the Relina luciferase activity. Data are means ± SEM of three independent experiments each analyzed in duplicates. *P < 0.05. (E) Effect of miR-101 on TGFB1 promoter activity. Cultured HepG2 were transfected with 15 nM of miR-101 or scrambled control for 24 h. Cells were cotransfected with plasmids containing firefly luciferase fused to TGFB1 promoter and control Relina luciferase and incubated for additional 24 h. Cells were treated with DMSO or TPA (100 nM) for 6 h in a fresh culture medium before the luciferase activity assay. TGFB1 promoter activity was normalized to the Relina luciferase activity. Data are means ± SEM of three independent experiments each analyzed in duplicates. *P < 0.05.