Fig. 2.

miR-181d downregulates MGMT expression. (A) Transfection of miR-181d suppressed MGMT mRNA (top panel) and protein expression in the A1207 glioblastoma cell line (bottom panel). For qRT-PCR, MGMT mRNA was normalized either to β-actin mRNA (shown here) or U6 rRNA (with comparable results, data not shown). For the Western blotting, α-tubulin was used as the loading control. (B) miR-181d binding sites in the MGMT 3′UTR suppressed luciferase expression in the presence of miR-181d. (C) Predicted miR-181d binding sites in the MGMT 3′UTR. (D), miR-181d physically interacts with MGMT transcript. β-actin was used as an internal control.