Mgf c25e

The migration was assessed through a wound healing assay. The tenocytes were seeded at a density of 4 × 10⁴ cells/well in a 24-well culture plate. After reaching 80–85% confluence, the tenocytes were synchronized by serum starvation for 12 h. The confluent monolayer was then scratched with a 10-μL pipet tip and washed twice with PBS. The wells were filled with 1 mL of serum-free DMEM with or without MGF-C25E (Phoenix Pharmaceuticals, Burlingame, USA). To test whether chromatin decompaction inhibited the rate of cell migration, we examined the effect of the levels of DNA methylation on the rate of cell migration. The cells were incubated with histone deacetylase (HDAC) inhibitor trichostatin A (TSA) (Beyotime, Jiangsu, China) or methylase inhibitor 5’-deoxy-5’-methylthioadenosine (MTA) (Sigma-Aldrich, St. Louis, MO, USA) for 60 min at 37 °C before scratching. Images of the wounds were acquired at 0, 24, and 48 h through a microscope (Olympus, Japan). Using Image J, the levels of wound closure were assessed by calculating the ratio of the closure area to the initial wound area as follows:  × 100 [%], where Wn represents the percentage of wound closure, An represents the residual wound area at the metering time point (h), and A0 represents the initial wound area.