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. 2023 Apr 5;141(26):3184–3198. doi: 10.1182/blood.2022018619

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© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.

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Figure 2. — IL-34 triggered leukemic cell differentiation in vitro and in vivo. (A) Gene ontology enrichment analyses of upregulated and downregulated pathways after IL-34 treatment (100 ng/mL). (B) Heat map of top 30 differentially expressed genes after IL-34 treatment in C1498 cells. (C) Representative Wright-Giemsa–stained cytospins of BM cells from mice treated with IL-34 showing signs of granulocytic differentiation. Scale bars, 20 μm. (D-G) Flow cytometry analysis of mature myeloid cells in the BM of mice treated with vehicle, IL-34, or Ara-C (n = 6). Populations are gated on GFP⁺ cells (D) and CD45⁺CD33⁺ cells (E-G). (H-I) Flow cytometry analysis of CD11b⁺Gr-1⁺ population gated on GFP in the BM of IL-34 KO and WT mice with leukemia, treated with or without IL-34 (100 μg/kg; n = 8). (J) Wright-Giemsa–stained BM of mice from panel I. Scale bars, 20 μm. (K) Flow cytometry analysis of myeloid differentiation marker CD11b in different AML cell lines treated with IL-34 (100 ng/mL) or all-transretinoic acid (ATRA) (500 nM). (L) Flow cytometry analysis of CD11b and CD14 expression on viable myeloid (CD33⁺) cells from 17 primary AML samples before and after incubation with recombinant human IL-34 (rhIL-34). (M) Wright-Giemsa staining of AML cell lines and primary AML cells. Scale bars, 10 μm. Data are representative of at least 3 independent experiments with cohorts of the indicated number of mice per group. Statistical significance was calculated using one-way ANOVA with Tukey multiple comparison test.

Figure 2. — IL-34 triggered leukemic cell differentiation in vitro and in vivo. (A) Gene ontology enrichment analyses of upregulated and downregulated pathways after IL-34 treatment (100 ng/mL). (B) Heat map of top 30 differentially expressed genes after IL-34 treatment in C1498 cells. (C) Representative Wright-Giemsa–stained cytospins of BM cells from mice treated with IL-34 showing signs of granulocytic differentiation. Scale bars, 20 μm. (D-G) Flow cytometry analysis of mature myeloid cells in the BM of mice treated with vehicle, IL-34, or Ara-C (n = 6). Populations are gated on GFP⁺ cells (D) and CD45⁺CD33⁺ cells (E-G). (H-I) Flow cytometry analysis of CD11b⁺Gr-1⁺ population gated on GFP in the BM of IL-34 KO and WT mice with leukemia, treated with or without IL-34 (100 μg/kg; n = 8). (J) Wright-Giemsa–stained BM of mice from panel I. Scale bars, 20 μm. (K) Flow cytometry analysis of myeloid differentiation marker CD11b in different AML cell lines treated with IL-34 (100 ng/mL) or all-transretinoic acid (ATRA) (500 nM). (L) Flow cytometry analysis of CD11b and CD14 expression on viable myeloid (CD33⁺) cells from 17 primary AML samples before and after incubation with recombinant human IL-34 (rhIL-34). (M) Wright-Giemsa staining of AML cell lines and primary AML cells. Scale bars, 10 μm. Data are representative of at least 3 independent experiments with cohorts of the indicated number of mice per group. Statistical significance was calculated using one-way ANOVA with Tukey multiple comparison test.