This supplementary material provides comprehensive supporting data for the study "N-acetyltransferase NAT10 controls cell fates via connecting mRNA cytidine acetylation to chromatin signaling" by Zhensheng Hu et al. The materials include figures and tables that detail various aspects of the research: 1. **Fig. S1**: Characterization of NAT10 knockdown in hESCs, including RT-qPCR and Western blot analyses, immunofluorescence, and gene expression changes. 2. **Fig. S2**: Analysis of gene expression and histone modifications in NAT10 knockdown hESCs, including H3K4me3 and H3K27me3 levels. 3. **Fig. S3**: Role of NAT10 in lineage differentiation and cellular reprogramming, with immunofluorescence, RT-qPCR, and teratoma assays. 4. **Fig. S4**: Characterization of ac4C levels in poly(A) RNAs in hESCs. 5. **Fig. S5**: Further analysis of acRIP-seq data, including motif enrichment and GO enrichment. 6. **Fig. S6**: Regulation of ANP32B by NAT10 and its role in cell fate transitions, including reprogramming assays and RT-qPCR analysis. 7. **Fig. S7**: Networks of ANP32B interactome and chromatin-associated binding partners. 8. **Fig. S8**: ANP32B's role in gene expression in hESCs, including volcano plots and KEGG enrichment. 9. **Fig. S9**: Integrated analyses of multiple assays, including CUT&Tag signals and ATAC-seq data. 10. **Fig. S10**: Partial mediation of NAT10's effects on chromatin landscape by ANP32B. 11. **Fig. S11**: Graphical summary of the study. 12. **Table S1**: Key resources used in the study. 13. **Table S2**: Oligonucleotides used in the study. These supplementary materials provide detailed experimental data and analyses to support the main findings of the study.This supplementary material provides comprehensive supporting data for the study "N-acetyltransferase NAT10 controls cell fates via connecting mRNA cytidine acetylation to chromatin signaling" by Zhensheng Hu et al. The materials include figures and tables that detail various aspects of the research: 1. **Fig. S1**: Characterization of NAT10 knockdown in hESCs, including RT-qPCR and Western blot analyses, immunofluorescence, and gene expression changes. 2. **Fig. S2**: Analysis of gene expression and histone modifications in NAT10 knockdown hESCs, including H3K4me3 and H3K27me3 levels. 3. **Fig. S3**: Role of NAT10 in lineage differentiation and cellular reprogramming, with immunofluorescence, RT-qPCR, and teratoma assays. 4. **Fig. S4**: Characterization of ac4C levels in poly(A) RNAs in hESCs. 5. **Fig. S5**: Further analysis of acRIP-seq data, including motif enrichment and GO enrichment. 6. **Fig. S6**: Regulation of ANP32B by NAT10 and its role in cell fate transitions, including reprogramming assays and RT-qPCR analysis. 7. **Fig. S7**: Networks of ANP32B interactome and chromatin-associated binding partners. 8. **Fig. S8**: ANP32B's role in gene expression in hESCs, including volcano plots and KEGG enrichment. 9. **Fig. S9**: Integrated analyses of multiple assays, including CUT&Tag signals and ATAC-seq data. 10. **Fig. S10**: Partial mediation of NAT10's effects on chromatin landscape by ANP32B. 11. **Fig. S11**: Graphical summary of the study. 12. **Table S1**: Key resources used in the study. 13. **Table S2**: Oligonucleotides used in the study. These supplementary materials provide detailed experimental data and analyses to support the main findings of the study.