Figure 1. An age-dependent MPN/AML model driven by HG stress and preleukemic mutation.

(A) Schematic for the generation of compound mutant Tet2^+/– Ins2^Akita/+ mice along with WT, Tet2^+/–, and Ins2^Akita/+ controls. (B) Kaplan-Meier survival curve for the compound mutant Tet2^+/– Ins2^Akita/+ mice and controls. Only compound mutant Tet2^+/– Ins2^Akita/+ mice showed early mortality, between 6 and 9 months of age, with 100% penetration. The median survival for the compound mutant mice was approximately 7 months (221 days), whereas all control mice survived beyond 300 days, after which the experiment was discontinued. (C) At the end stage of life (6–9 months of age), all the compound mutant Tet2^+/– Ins2^Akita/+ mice displayed a visible myeloid sarcoma under the skin around the ocular area on 1 or both sides (red arrow), as well as pale bones (blue arrow) and splenomegaly. (D) Spleen weights at 6–9 months of age, when the compound mutant mice became moribund. (E) Mutant Ins2^Akita/+ and Tet2^+/– Ins2^Akita/+ mice manifested an increase in blood glucose levels and a decrease in body weight compared with WT and Tet2^+/– mice throughout all stages. The plots show data for the mice at 5 months of age. (F) Hematologic analysis of PB using the automatic blood cell counter Element HT5. NE, neutrophil. (G) Analysis of myeloid cell compartments by flow cytometry using staining with Ly6G and Mac1 antibodies. (H) Analysis of BM mononuclear cells by flow cytometry using antibodies that recognize cKit and Mac1. (I) Quantification of the frequency of HSPCs and mature cells in the BM. n = 4–10 mice. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA (D–I). Data in B were analyzed with GraphPad Prism 6, using the Kaplan-Meier survival package. Freq., frequency.