Effects of sgRNA targeting cyclin D1 on cell-cycle and apoptosis.

<p>(A) Real-time visualization of cell cycle progression using a fluorescence microscope. HSC-3 cells were plated and cultured for 24 h. <i>In vitro</i> visualization of cell cycle distribution in a fluorescent ubiquitination-based cell cycle indicator (FUCCI) was used. Naked sgH5, sgLucHep1 (unrelated heptamer) or vehicle (None) was added at 200 nM and then cell cycle distribution was monitored using a fluorescence microscope. Scale bars = 100 µm. (B) Cellular activities of caspases 3/7 were measured 6 h after HSC-3 cells were cultured in the absence (–) or presence of naked sgRNA (200 nM each). siRNA (20 nM) for cyclinD1 with Lipofectamine 2000 was used as positive control. Fold-increase in activity was calculated based on activity measured from control cells. (C) Caspase 3/7 activity of cells treated with sgRNA targeting cyclin D1. HSC-3 cells were plated and cultured for 24 h. Naked sgHT5, sgH5 or sgL5 was added at 200 nM and then caspase 3/7 activity was visualized by a caspase 3/7 Green Detection Reagent. Each assay represents a separate experiment performed in triplicate. Data are means ± S.D.*<i>P</i><0.05.