Figure 3.

Direct regulation of BCL6 expression by miR-10a at the posttranscriptional level. (A) Quantitative RT-PCR analysis of miR-10a levels in OCI-LY7 and OCI-LY3 cells treated with pre-miR-control, pre-miR-10a, anti-miR-control or anti-miR-10a. U6 snRNA was used as an internal control, and the relative amount of miRNA normalized to the U6 snRNA levels was calculated using the 2^-ΔΔCT formula, in which ΔΔC_T = (C_{T miRNA} - C_{T U6}) _target - (C_{T miRNA} - C_{T U6}) _control. (B and C) Western blot analysis of BCL6 protein levels in OCI-LY7 and OCI-LY3 cells treated with pre-miR-control, pre-miR-10a, anti-miR-control or anti-miR-10a. B: representative image. C: quantitative analysis. (D) Quantitative RT-PCR analysis of BCL6 mRNA levels in OCI-LY7 and OCI-LY3 cells treated with pre-miR-control, pre-miR-10a, anti-miR-control or anti-miR-10a. (E) Direct recognition of the BCL6 3′-UTR by miR-10a. HEK293T cells were co-transfected with firefly luciferase reporters containing either wild-type (WT) or mutant (MUT) miR-10a binding sites in the BCL6 3′-UTR and pre-miR-control, pre-miR-10a, anti-miR-control or anti-miR-10a, 24 h after transfection, the cells were assayed using a luciferase assay kit. Data are the mean±SEM of 3 independent experiments performed in triplicate, ** P < 0.01; *** P < 0.001