Figure 2.
Deletion of the 9p21s region causes MDS/MPN-like disease. (A) Kaplan-Meier plots of R26CreER9p21fl/fl mice (n = 20) and WT mice (n = 12). (B) Kaplan-Meier plots of 9p21+/− mice with and without Flt3 mutant (9p21+/− mice, n = 43; 9p21+/−Flt3+/ITD mice, n = 8; WT mice, n = 23). P value shows log-rank test result. (C) Representative pale bones and splenomegaly in 9p21+/− tumor mice compared with that in WT mice. (D) Quantification of the spleen and liver weight. Symbols represent individual mice (WT mice, n = 22; 9p21+/− tumor mice, n = 38). (E) Complete blood count analysis of 9p21+/− tumor mice and WT mice. Symbols represent individual mice (9p21+/− tumor mice, n = 15; WT mice, n = 17). (F) May-Grünwald-Giemsa staining of peripheral blood smear. Red arrows indicate representative immature myeloid cells; the orange arrow indicates dysplastic neutrophil; white stars indicate dysplastic red blood cells. (G-H) Hematoxylin and eosin staining of BM sections. Dotted orange circles and a dotted green circle indicate multinucleated and small binucleated megakaryocytes, respectively (G); orange arrows indicate accumulated hemosiderin in abnormal megakaryocytes (H). (I) Absolute numbers of hematopoietic cells in the BM and spleen were analyzed via flow cytometry. Symbols represent individual mice (WT mice, n = 24; 9p21+/− tumor mice; n = 24). (J-L) The frequency of hematopoietic populations was analyzed via flow cytometry. (J) The frequency of long-term HSC (LT-HSC) in mice BM and spleen. Symbols represent individual mice BM (WT mice, n = 14; 9p21+/− tumor mice, n = 13) and spleens (WT mice, n = 10; 9p21+/− tumor mice, n = 11). (K) The frequencies of hematopoietic progenitors in mice spleens. Symbols represent individual mice. LSK (WT mice, n = 14; 9p21+/− tumor mice, n = 13); MPP (WT mice, n = 11; 9p21+/− tumor mice, n = 9); MkP (WT mice, n = 9; 9p21+/− tumor mice, n = 7); and EryP (WT mice, n = 5; 9p21+/− tumor mice, n = 5). (L) The frequencies of hematopoietic lineages in the spleens of WT and 9p21+/− tumor mice. Symbols represent individual mice. WT mice, n = 11; 9p21+/− tumor mice, n = 9. (M) Schematic representation of 9p21+/− MDS/MPN mouse BM and spleen cells transplantation experiment approach. (N) Kaplan-Meier plots of 9p21+/− MDS/MPN mouse BM and spleen cell–derived recipients. WT BM–derived recipients (lethal irradiation, n = 5); 9p21+/− donor 1 BM–derived recipients (lethal irradiation, n = 6), and spleen cell–derived recipients (lethal irradiation, n = 6); and 9p21+/− donor 2 spleen–derived recipients (sublethal irradiation, n = 6). Data represent the mean ± SD. Statistical significance was defined using Mann-Whitney test and is shown as follows: ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001 in panels D-E,I-L. EryP, erythroid progenitors; Gran, granulocyte; LSK, lineage− Sca1+cKit+ population; MPP, multipotent progenitors; Mono, monocyte.

Deletion of the 9p21s region causes MDS/MPN-like disease. (A) Kaplan-Meier plots of R26CreER9p21fl/fl mice (n = 20) and WT mice (n = 12). (B) Kaplan-Meier plots of 9p21+/− mice with and without Flt3 mutant (9p21+/− mice, n = 43; 9p21+/−Flt3+/ITD mice, n = 8; WT mice, n = 23). P value shows log-rank test result. (C) Representative pale bones and splenomegaly in 9p21+/− tumor mice compared with that in WT mice. (D) Quantification of the spleen and liver weight. Symbols represent individual mice (WT mice, n = 22; 9p21+/− tumor mice, n = 38). (E) Complete blood count analysis of 9p21+/− tumor mice and WT mice. Symbols represent individual mice (9p21+/− tumor mice, n = 15; WT mice, n = 17). (F) May-Grünwald-Giemsa staining of peripheral blood smear. Red arrows indicate representative immature myeloid cells; the orange arrow indicates dysplastic neutrophil; white stars indicate dysplastic red blood cells. (G-H) Hematoxylin and eosin staining of BM sections. Dotted orange circles and a dotted green circle indicate multinucleated and small binucleated megakaryocytes, respectively (G); orange arrows indicate accumulated hemosiderin in abnormal megakaryocytes (H). (I) Absolute numbers of hematopoietic cells in the BM and spleen were analyzed via flow cytometry. Symbols represent individual mice (WT mice, n = 24; 9p21+/− tumor mice; n = 24). (J-L) The frequency of hematopoietic populations was analyzed via flow cytometry. (J) The frequency of long-term HSC (LT-HSC) in mice BM and spleen. Symbols represent individual mice BM (WT mice, n = 14; 9p21+/− tumor mice, n = 13) and spleens (WT mice, n = 10; 9p21+/− tumor mice, n = 11). (K) The frequencies of hematopoietic progenitors in mice spleens. Symbols represent individual mice. LSK (WT mice, n = 14; 9p21+/− tumor mice, n = 13); MPP (WT mice, n = 11; 9p21+/− tumor mice, n = 9); MkP (WT mice, n = 9; 9p21+/− tumor mice, n = 7); and EryP (WT mice, n = 5; 9p21+/− tumor mice, n = 5). (L) The frequencies of hematopoietic lineages in the spleens of WT and 9p21+/− tumor mice. Symbols represent individual mice. WT mice, n = 11; 9p21+/− tumor mice, n = 9. (M) Schematic representation of 9p21+/− MDS/MPN mouse BM and spleen cells transplantation experiment approach. (N) Kaplan-Meier plots of 9p21+/− MDS/MPN mouse BM and spleen cell–derived recipients. WT BM–derived recipients (lethal irradiation, n = 5); 9p21+/− donor 1 BM–derived recipients (lethal irradiation, n = 6), and spleen cell–derived recipients (lethal irradiation, n = 6); and 9p21+/− donor 2 spleen–derived recipients (sublethal irradiation, n = 6). Data represent the mean ± SD. Statistical significance was defined using Mann-Whitney test and is shown as follows: ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001 in panels D-E,I-L. EryP, erythroid progenitors; Gran, granulocyte; LSK, lineage− Sca1+cKit+ population; MPP, multipotent progenitors; Mono, monocyte.

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