Fig. 2

miR-101-3p induces apoptosis in HCT116 cells. Cells were transfected with control miRNA (PMC), miR-101-3p or a control lethal siRNA against ubiquitins (siUBC), cultured for 72 h in 96-well plates, stained for apoptotic markers, and submitted to quantitative automated fluorescence microscopy. (a) Representative images of HCT116 cells stained with Annexin V (red), Propidium Iodide (PI, yellow) and Hoechst (blue). (b) Quantification of apoptotic markers separated cells into four populations: viable (Hoechst only), early apoptotic cells (double positive for Annexin V and Hoechst only), late apoptotic (triple positive for PI, Annexin V, and Hoechst), and necrotic (double positive for PI and Hoechst only). Quantification plots show significant increase in necrotic and late apoptotic cells with decrease in viable cells after treatment with miR-101-3p in comparison to control PMC (P < 0.05, unpaired one-tailed non-parametric Mann-Whitney test; n = 5 replica wells, 9 sites/well)