Histone H3 acetylation (pan-acetyl) of WT, Mpc2-deficient basal cells and acetate treated Mpc2-deficient basal cells

To further access whether changes in histone acetylation are directly associated with cell differentiation, we performed CUT&Tag experiment on WT cells, Mpc2-deficient cells, and acetate treated Mpc2-deficient cells by using antibody against H3 pan-acetylation. Mpc2 mutant cells displayed a decrease in the peaks of histone H3 acetylation near transcription start sites compared to control cells, whereas this reduction could be partially reversed by acetate supplementation. Pathway analysis using differentially acetylated genes between control and Mpc2 mutant cells revealed alterations in differentiation-related signaling pathways, such as TGF-ß and Notch. Indeed, Mpc2 mutant BCs showed insufficient H3 acetylation at the promoters of key differentiation related genes, such as Hes1 and HeyL, downstream transcriptional factor of Notch signaling, and the decrease could be partially reversed by acetate treatment. Overall design: Primary airway basal cells were isolated from WT and p63CreER; Mpc2f/f mice, followed by ALI culture. WT, Mpc2-deficient and acetate treated Mpc2-deficient cells were collected on day 3 of ALI culture for analysis.