Simultaneous lack of β- and γ-catenin in HSCs of mixed BM chimeras and serial transplants does not affect their repopulation capacity, nor does it influence B- or T-cell development. (A) Mixed BM chimeras were analyzed between 6 and 26 weeks after reconstitution with a 1:2 mixture of CD45.1⁺ and either control or β/γ dko CD45.2⁺ BM cells (left panel). BM cells from primary chimeric mice of either control or β/γ dko were serially transplanted into CD45.1⁺ recipients and analyzed at 12 and 24 weeks after reconstitution (right panel). Percentages of CD45.2⁺ KLS (CD117⁺lin⁻Sca1⁺), LT-HSCs (CD117⁺lin⁻Sca1⁺CD34⁻CD135⁻), ST-HSCs (CD117⁺lin⁻Sca1⁺CD34⁺CD135⁻), and MPPs (CD117⁺lin⁻Sca1⁺CD34⁺CD135⁺) from either control (□) or β/γ dko (■) mixed BM chimeras (n = 6 for control and n = 7 for β/γ dko) and the serial transplants (n = 5 for control and n = 5 for β/γ dko) are shown. (B) Percentages of myeloid lineages, including granulocytes (Gr; Gr1⁺Mac1⁺), macrophages (MØ; Gr1⁻Mac1⁺), megakaryocytes (Mega; CD41⁺), early erythroblasts (EB; Ter119⁺CD71⁺), and red blood cells (RBC; Ter119⁺CD71⁻) gated on CD45.2⁺ donor-derived cells derived from either control (□) or β/γ dko (■) mixed BM chimeras (n = 6 for control and n = 7 for β/γ dko). (C) Percentages of BM-derived B-cell subsets, including total BM B cells (B220⁺), pro-/pre-B cells (B220⁺CD43⁺BP1⁻), large pre-BII cells (preBII; B220⁺CD43⁺BP1⁺), small pre-BII cells (spreBII; B220⁺CD43⁻BP1⁺IgM⁻), and immature B cells (Imm; B220⁺CD43⁻BP1⁻IgM⁺), gated on CD45.2⁺ donor-derived cells derived from either control (□) or β/γ dko (■) mixed BM chimeras (n = 6 for control and n = 7 for β/γ dko). (D) Representative FACS analysis of CD45.2⁺ total thymocytes stained with anti-CD4 and anti-CD8 antibodies (top row), or with anti-CD44 and anti-CD25 antibodies (bottom row) after gating on Lin⁻ cells (and excluding CD44⁺CD117⁻ cells) derived from control and β/γ dko mixed chimeras. (E) Bar graphs represent relative percentages of CD45.2⁺ thymocyte subsets; DN (double negative; CD4⁻CD8⁻TCRβ⁻), DN1 (CD117⁺CD44⁺CD25⁻), DN2 (CD117⁺CD44⁺CD25⁺), DN3 (CD44⁻CD25⁺), DN4 (CD44⁻CD25⁻), DP (double positive; CD4⁺CD8⁺), CD4 (CD4⁺CD8⁻), and CD8 (CD4⁻CD8⁺) derived from control (□) and β/γ dko (■) mixed chimeras (n = 6 for control and n = 7 for β/γ dko). The error bars represent mean (± SD) in panels A, B, C, and E. No statistically significant differences (P values ranged between .06 and .89) were observed between control and β/γ dko chimeras in all populations analyzed.