Figure 2.
miR-197 depletion impairs cell proliferation and induces apoptosis. (a) Growth curve of NIH-H460 cells untreated (nt), transfected with control LNA or LNA-197 at 25 nM; cell number was assessed by Cell Titer-Glo assay at the indicated time points after transfection – mean±S.D., *P=0.034, **P<0.001. (b) Clonogenic capacity of NIH-H460 cells after anti-miR-197 treatment; the graph shows the percentage of plated cells that gave rise to colonies – mean±S.D., ***P<0.0001. (c) Activation of the apoptotic pathway after LNA-197 transfection is shown by CASPASE 3–7 activity, Annexin V staining (d) and immunoblot using the specified antibodies at the indicated time points (e) – mean±S.D., *P=0.04, **P=0.0086, ***P<0.0005. (f) NIH-H460 cells were transfected in vitro with control LNA or LNA-197; 16 h after transfection, 105 viable cells were injected into the flank of CD1/nude mice. Shown is the tumor growth of xenografts as defined by mass volume – mean±S.E.M., N=8. In the LNA-197-treated group, the volume of the only three measurable tumors was graphed. (g) Comparison of tumor engraftment sizes of LNA-197-treated versus control LNA-treated NIH-H460 cells. Tumors explanted from three mice 32 days after injection are shown