Figure 2

Functional effects of miR-10a. A. Transfection of OCI-AML3 cells with anti-miR10a LNA resulted in 19% reduction in cell count at 48 h as determined by MTS assay. B Assessment of clonogenic potential found a significant decrease in mean colony number (left), and a trend towards reduced Colony Size (right). Values are normalised to Scrambled LNA Control. C A significant increase in the number of Annexin-V positive events at 24 and 48 h. D Cell cycle analysis demonstrated an increase in the proportion of cells in the SubG1 fraction at 24 and 48 h post miR-10a knockdown. Black bars = Scrambled LNA Control, White bars = Anti-miR10a LNA transfected OCI-AML3 cells. E Confirmation of predicted miR-10a gene targets by luciferase reporter assay: ARNT, GTFH1, ID4, KLF4, MAPRE1, NR4A3, RB1CC1 and TFAP2C were confirmed to be miR-10a repressible by this assay. White bars = miR-10a repression of 3'UTR of the gene of interest, Black bars = miR10a repression of mutated miR-10a binding site on the 3'UTR of the gene of interest. All values were subject to normalisation to renilla luciferase activity of a non-specific transfection control. Error bars denote SEM. For all experiments n ≥ 3, statistical significance determined by Two-Tailed T-Test. *p < 0.05, **p < 0.01, *** p < 0.001.