Whole-cell lysates were prepared and analyzed by immunoblotting as described (51). For in vitro kinase reactions, recombinant p38α and ΔNp63α proteins fused with glutathione S-transferase (SignalChem) were incubated in a reaction buffer containing [γ-32P]ATP at 30°C for 30 min and analyzed by SDS-PAGE and autoradiography. To map phosphorylated ΔNp63α residues, SDS-PAGE–separated and Coomassie Blue–stained proteins were subjected to trypsin or chymotrypsin digestion and mass spectrometry [liquid chromatography– tandem mass spectrometry (LC-MS/MS); the BIDMC Mass Spectrometry Facility]. Total RNA was isolated using the RNeasy Mini Kit (Qiagen) and analyzed by real-time qPCR using gene-specific primers (table S2). DNA microarray analysis was performed using GeneChip Mouse Genome 430 2.0 Array (Affymetrix) and the GENE-E matrix visualization and analysis platform (Broad Institute).

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