Apoptosis assay
Apoptotic cells in NC, miR-210 mimic and miR-210 inhibitor groups were
quantified
using
the Annexin V-FITC apoptosis detection
kit (Beyotime Institute of
Biotechnology, Jiangsu, China). Both the attached cells and cells in the
supernatant were collected from the above four groups and fixed with 2%
paraformaldehyde. They were washed twice with cold phosphate-buffered
saline (PBS) and resuspended in 100 μl of the annexin-binding buffer.
Then, H9C2 cells were incubated with 2.5 μl of Annexin V for 15 min and
analysed using an EPICS XL flow cytometer (Beckman-Coulter, Fullerton,
USA). Apoptotic cells were defined as the cells that belonged to the
right quadrant (right top + right bottom) in flow cytometry plots. The
experiment was independently repeated three times, and an average
apoptotic rate was calculated.