Fig. 2
The effects of ACE on cytotoxicity and proliferation of HaCaT cells. ACE dehydrate was dissolved in sterilized water in a concentration of 50 mg/ml before use. (A) Cells (5 × 103) were seeded on a 24-well plate and incubated overnight at 37 °C with 5% CO2, and different concentrations of ACE or vehicle agent were added followed by incubation for 24–48 h. The MTT assay was conducted to determine the effect of ACE on cell viability. (B) The cell viability was also determined by flow cytometry with Annexin V-FITC/PI staining after the treatment with 5 mg/ml ACE for 48 h. (C) Cells were seeded on a 12-well plate and treated with different concentrations of ACE. The live cells were counted by trypan blue staining every 24 h. (D) Cells seeded on a 96-well plate were starved for 24 h, then cultured with fresh DMEM containing 10% FBS and treated with indicated concentrations of ACE. Cells were collected after 48 h-incubation to determine BrdU signal intensity according to the manufacturer’s protocol. * p < 0.05 compared to vehicle-treated cells.
