Fig. 2

Deletion of Fbxo22 fails to affect normal hematopoiesis. A Schematic of the Fbxo22 floxed allele showing the deletion of floxed exons 3–4 following Cre recombinase activity. Use of Mx1-Cre or Scl-Cre-ER^T results in specific deletion in HSCs following pIpC or tamoxifen treatment. B Fbxo22 deletion was evaluated by genotyping in PBMC and BM at indicated times after pIpC treatment (left), qRT-PCR (middle) and Western blot (right) in total BM cells at day 21 after pIpC treatment in mice. C, D Representative flow cytometry plots (left) and the percentages of LSK cells (Lin⁻Sca-1⁺c-Kit⁺), LT-HSCs (Lin⁻Sca-1⁺c-Kit⁺CD150⁺CD48⁻), MPP1 (Lin⁻Sca-1⁺c-Kit⁺CD150⁻CD48⁻), MPP2 (Lin⁻Sca-1⁺c-Kit⁺CD150⁺CD48⁺), and MPP3 (Lin⁻Sca-1⁺c-Kit⁺CD150⁻CD48⁺) (C, right, n = 5) and LK cells (Lin⁻Sca-1⁻c-Kit⁺), CMPs (Lin⁻Sca-1⁻c-Kit⁺CD16/32⁻ CD34⁺), GMPs (Lin⁻Sca-1⁻c-Kit⁺CD16/32⁺CD34⁺) and MEPs (Lin⁻Sca-1⁻c-Kit⁺ CD16/32⁻CD34⁻) in BM from Fbxo22^+/+ and Fbxo22^−/− mice (D, right, n = 5). (E–K) Primary competitive transplantation assay was conducted with Fbxo22^+/+ and Fbxo22^−/− BM CD45.2 cells (1 × 10⁶) along with CD45.1 competitor cells (1 × 10⁶). Percentages of donor or recipient-derived PB cells at the indicated times (E), and the indicated cells were analyzed at 16 weeks (F–K, n = 5). L Primary transplantation was conducted with sorted LT-HSCs (1 × 10³) from Fbxo22^+/+ and Fbxo22^−/− BM cells along with 1 × 10⁵ BM cells from CD45.1 competitor. Percentages of donor or recipient-derived cells in PB were analyzed at the indicated time points. Error bars denote mean ± SD. Statistical significance was determined by two-tailed unpaired t test. All animal experiments were repeated at least twice with similar results