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Fig. 5 Impaired engraftment ability of hematopoietic cells from adult WKM of csf1rb-deficient fish. (A) Work flow of WKM transplantation. Single-cell-stage embryos from the runx1w84x mutant were injected with pu.1 MO to create immunodeficient fish (runx1w84x; pu.1 morphant). The WKM cell suspension was prepared from csf1rb siblings and mutants with the Tg(coro1a:DsRedx) and Tg(globin:GFP) background and was transplanted into the vessel around the yolk of 2 dpf immunodeficient fish. These fish were raised to adults, and donor-derived blood cells were checked in the kidney, skin and circulation. Red circles indicate the coro1a-Red+ cells, green circles indicate the globin-GFP+ cells, the orange bar indicates the kidney, the black bar indicates the skin and the blue bar indicates the tail circulation. (B) Coro1a-Red+ cells on the skin of 3-month-old immunodeficient recipient fish after transplantation. Four out of 13 recipient fishes transplanted with WKM of sibling donor fishes were reconstituted by donor coro1a-Red+ cells on the skin (white arrows), but none of the 24 recipient fishes transplanted with WKM from csf1rb mutant donor fish had coro1a-Red+ cells on the skin. Scale bars: 200 μm. (C) Globin-GFP+ cells in the circulation in tails of 3-month-old immunodeficient recipient fish after transplantation. The four fish transplanted with WKM of sibling donors with coro1a-Red+ cells on the skin were also reconstituted by donor-derived red blood cells in the circulation (white arrows), and none of the fishes transplanted with WKM from csf1rb mutant donors had donor-derived globin-GFP+ cells in the circulation. Scale bars: 200 μm. (D) Flow cytometry analysis of WKM of 3-month-old immunodeficient recipient fish after transplantation. The four fish transplanted with WKM of sibling donors with coro1a Red+ cells on the skin were also reconstituted by donor-derived coro1a-Red+ cells in the kidney marrow, and none of the fishes transplanted with WKM of csf1rb mutant donors had donor-derived coro1a-Red+ cells in the kidney. (E) Percentage of coro1a+ cells in the WKM of recipients transplanted with the WKM cells from csf1rb siblings (n=13) or mutants (n=24). Data are presented as mean±s.e.m.