Replication stress, chromatin hyper-compaction, and DEK accumulation exist in CD34 ⁺ cells from FA patients. (A–C) Representative flow plots of p-ATR, p-CHK1, and γH2AX staining in BM CD34⁺ cells from five healthy donors and six patients with FA. The bar graphs show the MFI of p-ATR p-CHK1, and γH2AX (n = 5–6 samples per group). (D) Representative heatmaps and line plots show ATAC peaks ± 5 kb from TSS in BM CD34⁺ cells from five healthy donors and five patients with FA. (E) Statistics and quantification of ATAC signal in BM CD34⁺ (n = 5 samples per group). (F) qRT-PCR analysis and the bar graph shows the expression of DEK gene in BM CD34⁺ cells from indicated human samples (n = 5–6 samples per group). (G) Representative western blot shows the expression of DEK and tubulin in BM CD34⁺ cells from five healthy donors and five patients with FA. (H) The violin plots show DEK expression in human HSC-containing cluster by analyzing published scRNA-seq data (GSE157591, five healthy donors and seven patients with FA). (I) BM CD34⁺ cells were enriched from three healthy donors. The cells were transduced with indicated shRNA lentivirus, followed by sorting EGFP⁺ CD34^+–transduced cells and further qRT-PCR analysis. The bar graphs show the expression of the indicated gene in transduced cells under indicated conditions. The gene expression was normalized to that of shControl (n = 3 technical replicates per group). The data presented in panels A–G are representative of two independent experiments. The results shown in panel E are illustrated as violin plots, and panels A–C and F are shown as mean ± SD. Panel I is represented as the mean value. Statistical analyses were performed using a two-tailed unpaired Student’s t test for panels A–F and a Wilcoxon test for panel H. P values are presented as follows: **P < 0.01; ***P < 0.001. Source data are available for this figure: SourceData F3.