Figure 4: Cell-autonomous development of transplantable myeloid leukaemia in Tet2/3 DKO mice.
From: Acute loss of TET function results in aggressive myeloid cancer in mice
(a) Competitive repopulation assay using bone marrow cells from inducible chimeric mice. Red blood cell-depleted CD45.2+ bone marrow cells from Tet2+/+ Tet3fl/fl Mx1-Cre− (WT) or Tet2−/− Tet3fl/fl Mx1-Cre+ (DKO) mice were mixed with equal numbers of CD45.1+ competitor cells and transplanted into lethally irradiated CD45.1+ congenic B6.SJL mice (see Supplementary Fig. 12a). At 4 weeks after transplantation, Tet3 deletion was induced by administration of pIpC and peripheral blood was examined for donor chimerism at the indicated time points after pIpC injection. (b) Analysis of multilineage differentiation as percentage of peripheral blood cells within donor-derived (CD45.2+) cells. Myeloid cells (Mac-1+), B cells (B220+), T cells (CD3ɛ+). (c) Kaplan–Meier curve representing percent survival of bone marrow chimeric mice that received WT or Tet2/3 DKO cells after pIpC injection (n=8 per group). No lethality was observed for recipients of WT bone marrow cells. (d) Kaplan–Meier curve representing percent survival of recipient mice transplanted with 2 × 106 splenocytes from WT and diseased Tet2/3 DKO mice (n=9 per group). (e) May–Grünwald–Giemsa-stained peripheral blood smears of recipient mice. Scale bar, 20 μm. (f,g) Enlargement of spleens and livers of recipient mice. Representative photographs of spleen (f) and liver (g) from recipients of control or Tet2/3 DKO splenocytes. Weights of spleen or liver are shown below (n=7 per group). Means±s.e.m. are shown. ***P<0.0005 (Student's t-test). (h) Representative photographs of femurs and tibiae from recipients of WT or Tet2/3 DKO splenocytes. (i) A representative flow cytometric analysis of myeloid-lineage cells (Gr-1+/Mac-1+) in bone marrow, spleen and blood of recipient mice (n=5 per each group). (j) A representative flow cytometric analysis of erythroid-lineage cells (Ter-119+/CD71+) in bone marrow and spleen of recipient mice (n=5 per each group). (k) Haematoxylin and eosin staining and immunohistochemistry (IHC) of livers with anti-myeloperoxidase show loss of normal liver structure and infiltration with myeloid cells. Top, × 4 magnification; middle and bottom, × 40 magnification. Scale bar, 200 μm (upper panel) and 60 μm (middle panel).
