2.4. Wound-Healing Assay

MDA-MB 231 and B16-F10 cells were harvested from the exponential growth phase, washed twice by 1× Phosphate Buffered Saline (PBS) and resuspended in the respective medium, seeded at the density of 10⁶ cells per well into a six-multiwell plate and incubated at 37 °C in a humidified atmosphere of 5% CO₂ for 24 h. Then, in each well, was made an horizontal slit with a white tip, in a central position; after a PBS 1× washing cells were incubated in the absence or presence of DMSO Ganoderma lucidum extract at different concentrations for 48 h. Cell migration on the slit of the confluent well was assessed at 0, 48 h, in each condition, by light microscopy reversed phase (Leica microsystems).