Invasion Assay of OSCC Cells

This assay was performed to examine the modulation of OSCC cells during invasion by treatment with PL metabolites. Briefly, the membranes of 6.5 nm transwell chambers with 8 µm pores (Corning, United States) were coated with 50ul of gelatinous matrix Miogel (2.4 mg/ml) with type I collagen (0.8 mg/ml) (Corning, Cat # 354236) (Salo et al., 2015 (link))and incubated for 12–16 h. Then, the cells previously treated and non-treated with PL were plated in 200ul of DMEM/F12 medium without FBS, and subjected to the invasion assay according to Rodrigues AN et al. (2022) (link) (Rodrigues et al., 2022 (link)). The invasion profile was assessed by measuring the absorbance of the cell dye at 650 nm using an Epoch plate reader (BioTek).