Fig 5.

MiR-145 binds to IRS1 3' UTR region and there were 2 binding sites. (A) 50 ng psiCHECK2 vector containing IRS13'UTR were co-transfected with different concentrations of miR-145 mimics (final concentrations were 0, 50, 100, 200 and 400 nmol/l) into 293T cells. 400 nmol/l non-targeting mimics control was also co-transfected with psiCHECK2 vector containing IRS13'UTR. Firefly luciferase activity was normalized to Renilla luciferase. Then fold changes were normalized to 0 concentration. Four replicates were made for each transfection, and the data were shown as mean ± SD. (B) Six putative binding sites of miR-145 in IRS1 3' UTR region were predicted. (C) -(H) Determination of miR-145 binding sites in the 3' UTR of IRS1. 50 ng psiCHECK2 vector containing binding sites or binding sites mutant at seed region were co-transfected with miR-145 mimics or non-targeting mimics control (final concentration was 400 nmol/l) into 293T cells. Firefly luciferase activity was normalized to Renilla luciferase. Then fold changes were normalized to psiCHECK2. Replicates and data analysis were the same as A. (C) Site 1, (D) Site 2, (E) Site 3, (F) Site 4, (G) Site 5, (H) Site 6.