2.9. Wound Scratch Assay

A seeding density of 2 × 10⁵ cells was planted in a 24-well plate, using 500 µl per well, and cultured 24 hours. When the cell growth and confluence were above 90%, a vertical scratch was made with the tip of a 200 µl pipette tip and rinsed three times with PBS. The cell debris was subsequently removed, and the serum-free medium changed. Alloimperatorin (150 µM) and control medium (0.1% DMSO) were added to the control and treatment group, respectively. They were then placed in a 37°C incubator for 24 hours. A high content analysis system (Perkin Elmer) was used to observe and take pictures of scratch wounds at 0, 6, and 24 hours. The experiment was repeated three times.