miR-16 impairs the colony formation ability of pancreatic tumor cells and down-regulates Bcl-2/FGF-2. The transfection of miR-16 mimic, inhibitor and the respective negative controls was carried out using Dharmafect2 transfection reagent as described in Materials and methods. (A) Clonogenic cell survival assay. Cells were washed 2 times with phosphate-buffered saline and seeded in triplicate (20,000 cells/10-cm dish) 24 h post-transfection. Cells were allowed to grow for an additional 2 weeks, followed by staining with 0.1% crystal violet for visualization and photography. Panel I, mimic negative control; panel II, miR-16 mimic oligonucleotide; panel III, inhibitor negative control; panel IV, miR-16 inhibitor. (B) Northern blot analysis. RNA was isolated from the negative control, miR-16 mimic- and inhibitor-transfected BxPC-3 cells and subjected to Northern blotting. Lane 1, mock; lane 2, mimic negative control; lane 3, miR-16 mimic; lane 4, inhibitor negative control; lane 5, miR-16 inhibitor; lane M, molecular weight markers. To normalize the hybridization, RNU48 probe was used as an internal control.