Fig. 4

Loss of Fbxo22 impairs the function of LSCs. A, B Flow cytometry plots (left) and the percentages of MKs (A) and LGMP (B) in BM from the primary transplanted recipients (right, n = 6). C, D Flow cytometry plots (left) and the percentages of MKs (C) and LGMP (D) in BM from the secondary transplanted recipients (right, n = 5). E Serial colonies and colony numbers (n = 3) formed by Fbxo22^+/+ and Fbxo22^−/− LGMP cells collected from the primary recipients. F Survival curves for recipients receiving Fbxo22^+/+ and Fbxo22^−/− LGMP cells upon secondary transplantation (n = 6). G Limiting dilution assays comparing the frequencies of LSCs in Fbxo22^+/+ and Fbxo22^−/− MLL-AF9⁺ BM cells. The indicated GFP⁺ Fbxo22^+/+ and Fbxo22^−/− MLL-AF9⁺ BM cells collected from primary recipients were co-transplanted with 2 × 10⁵ BM competitor cells into lethally irradiated recipients. The competitive repopulating units (CRUs) were calculated by L-Calc software. H, I Cell-cycle (H) or apoptosis (I) analysis of LGMP cells in BM from the secondary recipients transplanted with Fbxo22^+/+ and Fbxo22^−/− AML cells (n = 5–6). Error bars denote mean ± SD. Statistical significance was determined by two-tailed unpaired t test (A–E and H, I) or log-rank test (F), and the P values were shown. All animal experiments were repeated at least twice with similar results