Plasmid vectors expressing hemagglutinin-tagged p38α and p38αCA (gifts from D. Engelberg), MKK6EE (a gift from J. Han), and FLAG-tagged ΔNp63α (Addgene) were used in 293T cell transfection using Lipofectamine 2000 (Thermo Fisher Scientific). Lentiviral vectors expressing ΔNp63α and its variants were constructed using the backbone vector pCDH-CMV-MCS-EF1-copGFP (System Biosciences). To generate ΔNp63α variants harboring amino acid substitutions, mutations were introduced into the coding sequence using GeneArt Site-Directed Mutagenesis System (Thermo Fisher Scientific). Mouse p63-specific shRNA constructs (TRCN0000012749 and TRCN0000423330; Thermo Fisher Scientific) were among the RNAi Consortium clones. The human p63-specific shRNA construct was specific to 5′-acacacatggtatccagatgac-3′ in TP63. Lentiviral vectors expressing mouse and human p63-specific shRNA were based on pLKO.1. Lentiviral particles were generated by 293T cell transfection and quantified using the QuickTiter Lentivirus Titer Kit (Cell Biolabs). Keratinocytes were infected with lentiviruses in the presence of polybrene (8 μg/ml) for 4 hours and used for subsequent analyses 2 days later.

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