2.6. Skin Explant Cultures

Fresh mouse ears were split, cut into 1-cm² pieces, and floated dermal side down on 1 mL RPMI medium containing 10% FCS and Penicillin/Streptavidin in a 24-well plate at either RT (20–24 °C) or 32 °C for 24 h. Skin samples were either treated with tacrolimus (Prograf, Astellas Pharma, Tokyo, Japan) or SP600125 (Merck, Darmstadt, Germany) in the culture medium. Concentrations were used as indicated in the figure legends. After 24 h, media was taken, snap frozen in liquid nitrogen, and stored at −20 °C until further use. Media was analysed for cytokine expression via ELISA.