Figure 1.
Mutant IDH1 knockin mice suffer from anemia and dyshematopoiesis. (A) Complete blood count (CBC) analysis of peripheral blood from 3- to 5-month-old control (Ctrl) (VavCre/IDH1WT/WT) and IDH1-KI (VavCre/IDH1WT/R132Q) mice (n = 24 for Ctrl; n = 18 for IDH1-KI). (B) Macroscopic views of the lower limb skeleton (top) and limb BM (bottom) cell pellets from Ctrl and IDH1-KI mice. Results are representative of 8 pairs of mice. (C) Hematoxylin and eosin (H&E) staining (top) and immunostaining (bottom) to detect the erythroid-specific marker Ter-119 in sections of BM of lower limbs from Ctrl and IDH1-KI mice in panel B (n = 5/group). Scale bar, 100 μm. (D) Left, flow cytometric analysis of percentages of Ter-119+ and CD11b+ cells in nucleated BM cells from Ctrl and IDH1-KI mice. Right, quantitation of the data in the left panels. Data are the mean ± SEM (n = 3). (E) Macroscopic views (top) and H&E staining (bottom) of spleens from control and IDH1-KI mice (n = 5/group). Scale bar, 1 cm (top) or 100 μm (bottom). (F) H&E staining (top) and Ter-119 staining (bottom) of spleens from Ctrl and IDH1-KI mice (n = 3/group). Scale bars, 100 μm. (G) Left, flow cytometric analysis of percentages of Ter-119+ and CD11b+ cells among nucleated spleen cells from Ctrl and IDH1-KI mice. Right, quantitation of the results in the left panel. Data are the mean ± SEM (n = 3/group). (H) Serum erythropoietin (EPO) level analysis by ELISA from 3- to 5-month-old Ctrl and IDH1-KI mice. Data are the mean ± SEM (n = 6/group). H&E (I) and Perls’ Prussian Blue (J) staining of BM and spleen sections from Ctrl and IDH1-KI mice (n = 3/group). Blue arrowhead in panel I, deposited pigment. Scale bar, 50 μm (I) or 100 μm (J). (K-L) Left, flow cytometric analysis of cell death as determined by7-AAD/Annexin V staining of Ter-119+ erythroblasts (K) and CD11b+ myeloid cells (L) from the BM and spleen of Ctrl and IDH1-KI mice. Right, quantitation of percentages of live, apoptotic (7-AAD–Annexin V+), and necrotic (7-AAD+Annexin V+) cells in the left panels. Data are the mean ± SEM (n = 4). *P < .05; **P < .01; ***P < .001. RBC, red blood cells; HB, hemoglobin; HCT, hematocrit; MCV, mean corpuscular volume; n.s., not significant; PLT, platelet; WBC, white blood cells.

Mutant IDH1 knockin mice suffer from anemia and dyshematopoiesis. (A) Complete blood count (CBC) analysis of peripheral blood from 3- to 5-month-old control (Ctrl) (VavCre/IDH1WT/WT) and IDH1-KI (VavCre/IDH1WT/R132Q) mice (n = 24 for Ctrl; n = 18 for IDH1-KI). (B) Macroscopic views of the lower limb skeleton (top) and limb BM (bottom) cell pellets from Ctrl and IDH1-KI mice. Results are representative of 8 pairs of mice. (C) Hematoxylin and eosin (H&E) staining (top) and immunostaining (bottom) to detect the erythroid-specific marker Ter-119 in sections of BM of lower limbs from Ctrl and IDH1-KI mice in panel B (n = 5/group). Scale bar, 100 μm. (D) Left, flow cytometric analysis of percentages of Ter-119+ and CD11b+ cells in nucleated BM cells from Ctrl and IDH1-KI mice. Right, quantitation of the data in the left panels. Data are the mean ± SEM (n = 3). (E) Macroscopic views (top) and H&E staining (bottom) of spleens from control and IDH1-KI mice (n = 5/group). Scale bar, 1 cm (top) or 100 μm (bottom). (F) H&E staining (top) and Ter-119 staining (bottom) of spleens from Ctrl and IDH1-KI mice (n = 3/group). Scale bars, 100 μm. (G) Left, flow cytometric analysis of percentages of Ter-119+ and CD11b+ cells among nucleated spleen cells from Ctrl and IDH1-KI mice. Right, quantitation of the results in the left panel. Data are the mean ± SEM (n = 3/group). (H) Serum erythropoietin (EPO) level analysis by ELISA from 3- to 5-month-old Ctrl and IDH1-KI mice. Data are the mean ± SEM (n = 6/group). H&E (I) and Perls’ Prussian Blue (J) staining of BM and spleen sections from Ctrl and IDH1-KI mice (n = 3/group). Blue arrowhead in panel I, deposited pigment. Scale bar, 50 μm (I) or 100 μm (J). (K-L) Left, flow cytometric analysis of cell death as determined by7-AAD/Annexin V staining of Ter-119+ erythroblasts (K) and CD11b+ myeloid cells (L) from the BM and spleen of Ctrl and IDH1-KI mice. Right, quantitation of percentages of live, apoptotic (7-AAD–Annexin V+), and necrotic (7-AAD+Annexin V+) cells in the left panels. Data are the mean ± SEM (n = 4). *P < .05; **P < .01; ***P < .001. RBC, red blood cells; HB, hemoglobin; HCT, hematocrit; MCV, mean corpuscular volume; n.s., not significant; PLT, platelet; WBC, white blood cells.

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