Fig. 4: CDX2 is a target gene of KAT8-mediated H4K16ac.

a Venn diagram showing the overlap between KAT8 binding peaks (n = 25,944) and H4K16ac binding peaks (n = 21,324). b Genomic distribution of KAT8 and H4K16ac copeaks in mTSCs. c Venn diagram showing the overlap between genes with KAT8 and H4K16ac copeaks (n = 9339) and genes downregulated upon KAT8 loss (n = 1595). d Gene Ontology analysis for KAT8 and H4K16ac target genes (n = 528) by DAVID by Kappa Statistics (p < 0.05). e Heatmap of genes associated with stemness and proliferation that are the target genes of KAT8 and H4K16ac. f Genome browser view of CUT&Tag-seq signals and RNA-seq tracks for KAT8 and H4K16ac target gene Cdx2 in mTSCs. g Schematic of overexpressing CDX2 in Kat8^d/d mTSCs. The graphic elements were created by figdraw. h Immunoblot analysis of Flag and CDX2 in Kat8^f/f and Kat8^f/f transduced with 3xFLAG-CDX2 mTSCs. i Cell counts of Kat8^f/f, Kat8^d/d and Kat8^d/d + CDX2 mTSCs collected on indicated days. Data are representative of three independent experiments (n = 3). Two-tailed unpaired Student’s t-test. Error bars, mean ± SEM. P = 0.0463 (for D4, between Kat8^d/d and Kat8^d/d + CDX2), P = 0.0021 (for D5, between Kat8^d/d and Kat8^d/d + CDX2). Source data are provided as a Source Data file. j Immunofluorescent staining of CDX2, EOMES, pH3 in Kat8^f/f, Kat8^d/d and Kat8^d/d + CDX2 mTSCs. k Right panel: the relative fluorescence intensity of CDX2+ or EOMES+ signals in indicated groups, Data are representative of three independent experiments (n = 3) and the values are normalized to Kat8^f/f control group. Two-tailed unpaired Student’s t-test. Error bars, mean ± SEM. P = 0.003 (between Kat8^f/f and Kat8^d/d), P = 0.0026 (between Kat8^d/d and Kat8^d/d + CDX2). Left panel: the percentage of pH3⁺ cell numbers in mTSCs of indicated genotypes; average cell numbers from five 20X fields were determined. Data are representative of three independent experiments (n = 3) and the values are normalized to Kat8^f/f control group. Two-tailed unpaired Student’s t-test. Error bars, mean ± SEM. P = 0.009 (between Kat8^f/f and Kat8^d/d), P = 0.0176 (between Kat8^d/d and Kat8^d/d + CDX2). Source data are provided as a Source Data file. l Immunoblot analysis of CDX2, EOMES, KAT8 and H4K16ac in Kat8^f/f, Kat8^d/d and Kat8^d/d + CDX2 mTSCs. m Quantitative real-time PCR analysis of Elf5, Ly6a, and Eomes mRNA levels in Kat8^f/f, Kat8^d/d and Kat8^d/d + CDX2 mTSCs. Data are representative of three independent experiments (n = 3) and the values are normalized to ACTB and indicated as the mean ± SEM. Two-tailed unpaired Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.