2.7 Cell cycle assay
Each group of VSMCs was washed twice with ice-cold phosphate buffered
saline (PBS); cells were harvested by centrifugation and re-suspended in
PBS, then fixed and permeabilized with 70% ethanol. Cells were washed
twice in ice-cold PBS to remove ethanol, then treated with 500 μl
propidium iodide (PI; Wanleibio, China) in the dark at 4°C for 30 min.
Flow cytometry (ACEABio, NovoCyte, USA) was used to detect cell cycle.