4.7. Apoptosis Assays in Primary Keratinocytes

Apoptosis assays were performed as described [34]. Briefly, subconfluent 60 mm dishes of primary keratinocytes were treated for 15 h with 10 ng/mL TNF-α (Sigma-Aldrich, St Louis, MO, USA) and 10 µg/mL cycloheximide (Sigma-Aldrich, St Louis, MO, USA). Cells were trypsinized, and the percentages of viable cells were obtained by counting the cells in a Neubauer chamber. Experiments were performed 3 times per duplicate; in either experiment, pools from 3 to 6 animals of each phenotype were cultured together.