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tion, 4 lineages could be observed, erythroid cells, megakaryocytes, neutrophils, and macrophages, in the single SLAM-HSC generated clones after 10-14 days. First, Setd2Δ/Δ SLAM-HSCs showed decreased clonogenic- ities, as there were significantly less HSCs that could gen-
A
erate clones in each plate in the Setd2Δ/Δ group compared with the control, while the sorting efficiency was compa- rable between these two groups (Figure 4E). In addition, Setd2Δ/Δ SLAM-HSCs produced fewer frequencies of 4-lin- eage clones compared with the control, accompanied
B
CD
Figure 2. Setd2Δ/Δ mice showed profound reduction of myeloid, lymphoid and megakaryocyte progenitors but significantly increased erythroid progenitors. (A) Flow cytometry analysis of Setd2f/f and Setd2f/f/Vav1-Cre mice bone marrow (BM) cells. (B) Absolute number of hematopoietic progenitor cell (HPC) populations. [Setd2f/f =4, Setd2f/f/Vav1-Cre=5; mean±Standard Error of Mean (SEM)]. (C) Colony-forming cell (CFU) using BM cells from Setd2f/f or Setd2f/f/Vav1-Cre. 2x104 cells were plated in M3434 in triplicate and colonies were scored every seven days. GEMM: granulocyte, erythroid, macrophage, megakaryocyte colony; GM: granulocyte/macrophage; G/M: granulocyte or macrophage; BFU-E: burst formation unit-erythroid. Representative data were from 3 independent experiments. (N=3; mean±Standard Deviation (SD)]. (D) CFU-erythroid (CFU-E) assay using BM cells from Setd2f/f or Setd2f/f/Vav1-Cre. 5x105 cells were plated in M3334 in triplicate and colonies were scored 48 hours later. Representative data were from 3 independent experiments. (N=3; mean±SD).
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