Nr4a1/3-null HSCs have normal self-renewal, homing and engraftment capacity. (A) Serial replating of 100 sorted HSCs after acute Nr4a1/3 codepletion. From the second to the fifth round of replating, 10 000 cells were seeded. (B) LSK homing assay. CDKO and control mice (n = 3) were treated with 4 doses of tamoxifen, and LSK cells were sorted and subsequently labeled with CSFE in vitro. LSK CSFE⁺ cells were then transplanted into wild-type mice (n = 3 recipients per genotype) that were lethally irradiated the day before. Sixteen hours after transplant, recipient mice were killed and bone marrow was analyzed by flow cytometry for CSFE detection. Bone marrow from nontransplanted mice was used as negative control. (C) Flow cytometry analysis of CD45.2 donor cell chimerism in mice transplanted with 200 CDKO or control HSCs. Recipient mice were analyzed 6 weeks after transplant to confirm initial engraftment (Pre-TAM) and immediately treated with tamoxifen and analyzed every 4 weeks after tamoxifen treatment. (D) CD45.2 donor cell chimerism in the bone marrow at week 20. Results expressed as mean ± SD, n = 3 (A-B) or n = 3 control and 5 CDKO (C-D).