MiR-335 abrogation induces apoptosis in C6 astrocytoma cells. (A) Cell cycle distribution detection. C6 cells were transfected with 100 nM antagomir-335 or antagomir-NC for 72 h and cell cycle distribution was analyzed by flow cytometer. (B) Phase-contrast imaging and (C) Hoechest 33258 staining of C6 cells. (D) Quantitation of the percentage of apoptotic cells. (E, F) Apoptotic cells were detected by TUNEL assay. (G) Dose-dependent effect of antagomir-335 on caspase3/7 activity. C6 cells were transfected with antagomir-335 at the indicated concentrations for 72 h. Data represent the means ± SD. Statistical differences compared with the controls are given as *, P < 0.05; **, P < 0.01; ***, P < 0.001. Original magnification in (B, E), 200 ×; Original magnification in (C), 640 ×.