Fig. 1. miR-4779 inhibits tumor cell growth and induces apoptosis.

a–g HCT116 cells were transfected with miR-NC or miR-4779. At 72 h after transfection, cell viability was determined by MTS assay a. The non-transfected control (cont) was normalized to 100% cell viability and the effects of the miR-NC or miR-4779 were calculated accordingly. At 48 h after transfection, cell images were captured using phase contrast microscopy b. Magnification = ×100, scale bar = 100 μm. Cell cycle was analyzed by flow cytometry c and the percentage of cells in G1 and G2/M phase are annotated in each column. Apoptosis was measured by flow cytometric analysis of cells stained with Annexin V-FITC and PI d. The percentages of Annexin V-FITC only positive cells (right lower, early apoptosis) and both Annexin V-FITC and PI positive cells (right upper, late apoptosis) were quantitated. Right, the quantified apoptotic cell population was estimated. Equal amounts of cell lysates were subjected to western blotting using the indicated antibodies e. β-actin was used as a loading control. At 24 h after transfection, cells were re-plated and performed colony formation f and soft agar assays g. Each colony was stained, taken (left), and counted (right). Data are presented as averages of triplicate measurements with error bars representing standard deviations. **P < 0.01 and ***P < 0.001. All experiments were performed tree times on separate samples with comparable results