Figure 2.

miR-302d inhibits oxidant-induced cell death. hADSCs were transfected with miR-control or miR-302d for 48 h and were then treated with CoCl₂ or SIN-1. (a) Cells were treated with various concentrations of CoCl₂ or SIN-1 for 20 h and cell viability was assessed with trypan blue exclusion method. The results were presented as the percentage of total cells. (b) Flow-cytometric analysis of apoptotic cells. Cells were treated with 100 μM CoCl₂ or 5 mM SIN-1 for 24 h and the fraction of apoptotic cells displaying a subG1 DNA content was analyzed by FACS with PI. Cells with a sub-diploid DNA content (subG1) were considered to be apoptotic. (c) Analysis of apoptotic cells with Annexin V staining. Cells were treated with the indicated oxidant for 16 h. The fraction of apoptotic cells was analyzed by flow cytometry after the cells were stained with both Annexin V-FITC and PI. (d) Measurement of intracellular ROS generation using H₂-DCFDA staining. The negative control is unstained cells. ROS level is presented as the percentage of fluorescence intensity of the stained, control cells (right panel). *P<0.05, **P<0.01 versus CoCl₂ or SIN-1 treated miR-control. ^$P<0.05 versus untreated control. Data are shown as the mean±S.D. of three independent experiments