Fig. 2. Effect of harmine hydrochloride (HMH) on migration, invasion, and colony formation. (A) Cells after confluent growth in six-well plates were scratch-wounded and treated with 0~50 μM of HMH to measure the migration area after 24 h. (B) Cells were placed in the upper chamber with 1.5 mL of serum-free Dulbecco’s modified Eagle’s medium (DMEM), and DMEM containing 10% fetal bovine serum was added to the lower chamber. After culturing for 24 h, non-invading cells in the upper chamber were removed using a cotton swab, and cells invading the lower chamber were fixed using 4% paraformaldehyde and stained with 0.5% crystal violet solution. Subsequently, invasive cells were observed under a microscope. (C) Cells were seeded in six-well plates and treated with 0~50 μM of HMH for 24 h, and then, the medium was removed. Thereafter, cells were cultured for 14 days while replacing the medium with fresh medium every 3~4 days. Cells were fixed with methanol and stained with 0.5% crystal violet to determine colony formation. They were examined under a microscope, and wound areas and number of colonies were determined using ImageJ software. The results are presented as the mean±SD. Student’s t-test was used to analyze statistical differences between groups (*P<0.05, **P<0.01, ***P<0.001 with respect to the control group). Cell morphology was visualized using an inverted microscope at 200× magnification. 

Prev Nutr Food Sci 2023;28:436~443 https://doi.org/10.3746/pnf.2023.28.4.436