Supplementary Figure 5: Effect of absence of Flt3L and GATA-1 on myeloid progenitors.
From: Distinct myeloid progenitor–differentiation pathways identified through single-cell RNA sequencing
(a) Flow cytometry analysis of wild type and Flt3L KO pre-GM cells from 17 week old male mice, showing Flt3 expression. Numbers indicate mean percentage of cells in each gate (+/− SD). n=5 from 2 experiments. (b) Quantification of number of myeloid progenitors by flow cytometry analysis shown as percentage of total bone marrow. Number of cells of indicated progenitors are shown as frequency of total number of bone marrow cells (+/− SD). n=5 from 2 experiments. (c) Schematic overview of Gata1 KO studies. Chimeric mice with wild type and conditional Gata1 KO blood cells are created by transplanting indicated cells into irradiated recipients. pIC is injected to delete the floxed Gata1 gene mediated by MxCre. GE− and GE+ pre-GM and GMP cells were analyzed and sorted for in vitro culture. (d) Quantification of EGFP positive cells by flow cytometry of CD45.2 donor cells from experiment depicted in (c). Bars show percentage EGFP positive cells of the indicated CD45.2 progenitor population. Gata1fl/Y without MxCre are defined as wild type cells. n=2 for wild type cells and n=3 for Gata1 KO. SD is shown. (e) Indicated cells were manually plated at an average of 1 cell/culture and cultured for 8 days. Cell types of cultures are based on morphology analysis of cytospins. The number of cultures analyzed is indicated and are accumulated from 2 experiments. Mo: monocyte, PMN: polymorphonuclear granulocyte, Ma: mast cell (f) Representative picture of mast cells from cultured wild type and Gata1 KO GMP GE+ single cells. Scale bar is 25µM.
