2.6. Wound Healing Assay

Wound healing assay was used to assess the effects of alisol A on cell migration. MDA-MB-231 cells were seeded into 24-well plate (2 × 10⁵ cells/well) and cultured until confluent. The monolayer cells were scratched with a sterile 1000 μL pipette tip followed by washing with PBS to remove the floating cells. Cells were treated with alisol A for 24 h. Scratched areas were photographed (magnification 40×) at zero hour and then subsequently again 24 h later to assess the degree of wound healing. The migration rate was calculated according to the following equation: Wound closure % = 1 − (wound area at t₂₄/wound area at t₀) × 100%, where t₂₄ is the time after wounding and t₀ is the time immediately after wounding.