Il 36

The HaCaT cell line was obtained from Cell Lines Service (CLS Cell Lines Service, Germany; 300493) and maintained in Dulbecco's modified Eagle's medium (DMEM, high glucose, no glutamine) supplemented with 10% v/v fetal bovine serum (FBS), 1% v/v antibiotics (100 U/mL penicillin, 100 µg/ mL streptomycin), 1% v/v L-Glutamine at 37°C and 5% CO 2 in a humidified incubator. Cell culture reagents were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). Cells were passaged at 80% confluence. The culture medium was renewed every 3 days. For all experiments, HaCaT cells were incubated with exogenous cytokines: IL-6, IL-17A, IL-23, IL-36, TNF-α (all from Peprotech, USA) for 48 h at a final concentration of 100 ng/ml, or 1 µM imiquimod (IMQ, InVivoGene, USA) for 48 h [21] .