Inhibition of DEK restores the function of BM CD34 ⁺ cells from FA patients. (A) Experimental schematic. Freshly enriched human BM CD34⁺ cells were transduced twice with indicated shRNA lentivirus. Next, flow cytometry analysis and EdU incorporation assay were performed using the transduced cells (EGFP⁺CD34⁺). For transplantation, a total of 2 × 10⁴ EGFP⁺CD34⁺ cells were injected in the tail of sublethally irradiated (2.0 Gy) immunodeficient NSG mice. (B) qRT-PCR analysis and the bar graph shows DEK expression in EGFP⁺CD34⁺ cells (n = 3 human samples). (C) Representative flow plots of p-ATR staining in EGFP⁺CD34⁺ cells. The right bar graph shows the MFI of p-ATR and p-CHK1 (n = 3 human samples). (D) Representative flow plots show the EdU⁺ in EGFP⁺CD34⁺ cells. The right bar graph shows the EdU⁺ frequency (n = 3 human samples). (E) Representative flow plot of chimerism analysis in PB from recipient mice. (F) The left chart shows the donor (EGFP⁺hCD45⁺) chimerism in PB of the recipient mice at indicated time points after transplantation (n = 5 recipient mice per group). The right bar graph shows the donor chimerism in BM of the recipient mice at 16 wk after transplantation (n = 5 recipient mice per group). BM CD34⁺ cells from patient-2 were used for this experiment. shF, shFANCD2. (G) The donor chimerism in PB and BM of the recipient mice were analyzed (n = 5 recipient mice per group). BM CD34⁺ cells from patient-4 were used for this experiment. (H) Experimental schematic. BM CD34⁺ cells from healthy donors were transduced with shRNA lentivirus, followed by administrated with DEK aptamer (DTA-64) at the indicated concentration. After 72 h, cells were collected for analysis. (I) Representative flow plots show CD34⁺ cells. The right bar graph shows the CD34⁺ frequency (n = 3 samples per group). (J) Representative flow plots show γH2AX staining in BM CD34⁺ cells from two patients with FA after indicated aptamer treatment. (K) Representative flow plots show the EdU⁺ in BM CD34⁺ cells after indicated aptamer treatment, the right bar graphs show the EdU⁺ frequency (n = 3 samples per group). The data presented in panels B–K are representative of two independent experiments. Data are shown as mean ± SD. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post hoc test for panels B–K. P values are presented as follows: *P < 0.05, **P < 0.01, and ***P < 0.001; n.s., not significant.