HaCaT cells were seeded on 48-well plates (100,000 cells/well) a day prior to the experiment and stimulated with different concentrations of ursolic acid formulations in the presence of an M5 cytokin mix consisting of 1 ng/mL of each IL-17A, IL-22, oncostatin M, IL-1α, and TNF-α (BioLegend, BioCourse.pl, Katowice, Poland) diluted in DMEM medium. After 24 h, cell media were collected and stored at −20 °C for further analysis.
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