Supplementary materials for the study "N-acetyltransferase NAT10 controls cell fates via connecting mRNA cytidine acetylation to chromatin signaling" include figures S1 to S11 and tables S1 and S2. Figure S1 shows the generation and characterization of NAT10 knockdown human embryonic stem cells (hESCs), including gene expression analysis, knockdown efficiency, and effects on pluripotency markers. Figure S2 presents gene expression and histone modification analyses in NAT10 knockdown hESCs, highlighting changes in gene expression and histone marks. Figure S3 demonstrates NAT10's role in lineage differentiation and cellular reprogramming, including immunofluorescence, teratoma assays, and growth curves. Figure S4 characterizes ac4C levels in poly(A) RNAs from hESCs. Figure S5 provides further analysis of acRIP-seq data, including peak intersections and gene ontology enrichment. Figure S6 explores the role of ANP32B in NAT10-mediated cell fate transitions, including gene expression changes and chromatin remodeling. Figure S7 shows protein interaction networks of ANP32B. Figure S8 examines ANP32B's role in gene expression regulation in hESCs. Figure S9 integrates multiple assays to analyze chromatin landscape changes. Figure S10 illustrates ANP32B's partial mediation of NAT10 effects on chromatin. Figure S11 provides a graphical summary of the study. Table S1 lists key resources, including chemicals, peptides, and recombinant proteins. Table S2 provides oligonucleotides used in experiments. All figures and tables support the main findings of the study, showing the role of NAT10 in connecting mRNA acetylation to chromatin signaling and its impact on cell fate decisions.Supplementary materials for the study "N-acetyltransferase NAT10 controls cell fates via connecting mRNA cytidine acetylation to chromatin signaling" include figures S1 to S11 and tables S1 and S2. Figure S1 shows the generation and characterization of NAT10 knockdown human embryonic stem cells (hESCs), including gene expression analysis, knockdown efficiency, and effects on pluripotency markers. Figure S2 presents gene expression and histone modification analyses in NAT10 knockdown hESCs, highlighting changes in gene expression and histone marks. Figure S3 demonstrates NAT10's role in lineage differentiation and cellular reprogramming, including immunofluorescence, teratoma assays, and growth curves. Figure S4 characterizes ac4C levels in poly(A) RNAs from hESCs. Figure S5 provides further analysis of acRIP-seq data, including peak intersections and gene ontology enrichment. Figure S6 explores the role of ANP32B in NAT10-mediated cell fate transitions, including gene expression changes and chromatin remodeling. Figure S7 shows protein interaction networks of ANP32B. Figure S8 examines ANP32B's role in gene expression regulation in hESCs. Figure S9 integrates multiple assays to analyze chromatin landscape changes. Figure S10 illustrates ANP32B's partial mediation of NAT10 effects on chromatin. Figure S11 provides a graphical summary of the study. Table S1 lists key resources, including chemicals, peptides, and recombinant proteins. Table S2 provides oligonucleotides used in experiments. All figures and tables support the main findings of the study, showing the role of NAT10 in connecting mRNA acetylation to chromatin signaling and its impact on cell fate decisions.