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. 2021 Mar 23;218(5):e20201974. doi: 10.1084/jem.20201974

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© 2021 Chen et al.

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Figure S2. — DEK deletion decreases the HSPC pool in mice. (A) FACS analysis of CLPs (Lin⁻CD127⁺Sca-1^loc-Kit^lo) and LMPPs (Lin⁻ Sca-1⁺c-Kit⁺CD135⁺) in BM cells of Dek^fl/fl and Dek-cKO mice. (B and C) Count of CLPs and LMPPs in BM cells of Dek^fl/fl and Dek-cKO at 3 mo of age (n = 6–7). (D) Analysis of apoptotic HPCs and LSK cells in BM cells of Dek^fl/fl and Dek-cKO at 3 mo of age. Annexin V⁺ cells represent early and later apoptotic cells (n = 5). (E) Experimental schematic for the generation of mice with inducible deletion of DEK, Dek^fl/flMx1-Cre. Mice were treated with an i.p. injection of 10 µg pIpC per gram of body weight every second day for a total of three injections. The mice were sacrificed for analyses at 1 or 3 mo after pIpC injection. (F) Relative mRNA expression of DEK in freshly sorted HSCs of Dek^fl/fl and Dek^fl/flMx1-Cre mice at 1 mo after pIpC injection (n = 4). (G) FACS analysis of LSK cells and HPCs in Dek^fl/fl and Dek^fl/flMx1-Cre BM cells at 1 or 3 mo after pIpC injection. (H) BM cells count of Dek^fl/fl and Dek^fl/flMx1-Cre mice after pIpC injection (n = 5). (I) Count of HPC, CMP, GMP, and MEP in BM cells of Dek^fl/fl and Dek^fl/flMx1-Cre mice after pIpC injection (n = 5). (J) LSK cells count in Dek^fl/fl and Dek^fl/flMx1-Cre BM cells (n = 5). (K) HSC (CD48⁻CD150⁺LSK, CD34⁻LSK, or CD34⁻CD135⁻LSK) count in Dek^fl/fl and Dek^fl/flMx1-Cre BM cells (n = 5). (L) In vitro assay of the CFUs at 10–12 d after plating Dek^fl/fl and Dek-cKO HSC cells. For the second plating, live cells from the colonies obtained during the first plating were plated as before and cultured for 10–12 d (n = 5). Error bars represent means ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; Student’s t test. Data are representative of three independent experiments.