2.1.2. Scratch Wound Healing Migration Assay

Cell migration was assessed using a scratch wound healing assay following our previous publication [19]. In brief, the parental and CisR cells were cultured in six-well plates for 24 h and then treated with 50 µM cisplatin for another 24 h. Cells were re-suspended and again, 2 × 10⁵ cells were seeded into six-well plates and cultured to monolayers, which were then wounded using sterile 1 mL pipette tips. Cells were washed with PBS to remove any debris. Photos were captured at 0, 12, and 24 h after wounding (Figure 1). The gap distance can be quantitatively evaluated using software such as ImageJ (National Institutes of Health). The equations for calculation of the relative wound area (Equation (1)), percentage (%) of wound closure (Equation (2)), and wound healing speed (Equation (3)) are given below.

W₀ = Wound area at 0 h (µm²)

W_t = Wound area at ∆h (µm²)

ΔT = Duration of wound measured (h)

Representative images of scratch wound healing assay at 0, 12, and 24 h.