Fig. 2
Over-expression of miR-711 increased cell damage in response to oxidative stress. a Transfection with miR-711 mimics increased miR-711 expression. H9c2 cells were transfected with 1 μg miR-711 mimics, or control miRNA for 48 h. miR-711 expression was detected by RT-qPCR. Snord 61 was used as an internal loading control. Data were presented using the 2-∆∆CT method. b Over-expression of miR-711 increased cell apoptosis in response to AA. 48 h after transfection, H9c2 cells were treated with 3 h AA and 3 h reperfusion. Cell apoptosis/death was measured by Annexin-V and PI staining followed by flow cytometry. Left: Representative images of flow cytometric results; Right: Summarized data of flow cytometry from three independent experiments. c Inhibition of miR-711 reduced AA-induced cell apoptosis. H9c2 cells were transfected with miR-711 inhibitors 24 h prior to AA treatment. Cell death was detected as described in 2B. d miR-711 mimic promoted cell death induced by H/R, but miR-711 inhibitor decreased cell death, dynamically detected in an IncuCyte system. H9c2 cells were transfected with miR-711 mimic, mimic control, miR-711 inhibitor or inhibitor control for 48 h. Cells were then subjected to cold hypoxia (0.5% O2 at 10 °C) for 18 h. PBS was immediately replaced with DMEM-CM containing SYTOX Green dye and cells were then placed into an IncuCyte system for reoxygenation and cell death detection. Cells were scanned every hour. Images were taken at 20 h after reoxygenation (upper panel). The mean of green fluorescence per image was analyzed by the program provided by the IncuCyte system. A plot of green image means over reoxygenation time was generated (bottom panel). n = 3, * P < 0.05