Figure 3
Figure 3. Combined GM and lymphoid transcriptional priming in LSKFlt3hiMpl− BM cells. (A) Coexpression patterns of transcriptional lineage programs in single cells from BM LSK subpopulations. Cells were scored as expressing GM and/or lymphoid (Ly) programs based on the expression of one or more lineage-associated genes: GM: Csf3r and Mpo; lymphoid: Rag1, sterile IgH transcript, and Il7r. Mean plus or minus SEM values from 2 experiments with 88 cells investigated in each experiment. (B) Coexpression pattern of Mpl and Flt3 on Kit-enriched, lineage-negative (Lin−), Sca-1+ and Kit+ (LSK) fetal liver (day E14.5-E15.5) cells. Gates denote the sorting strategies used to purify LSKFlt3hiMpl− (Flt3hiMpl−), LSKFlt3hiMplhi (Flt3hiMplhi), and LSKFlt3− cells. Percentages indicate mean quadrant frequencies within LSK cells from 4 experiments. Panels below show typical purity analysis for LSKFlt3hiMpl− and LSKFlt3hiMplhi cells. (C) In vitro Mk potential of fetal liver LSKFlt3−, LSKFlt3hiMplhi, and LSKFlt3hiMpl− cells, investigated as described in “Methods” after 8 days of culture. Mean plus or minus SEM values from 4 experiments. (D) Coexpression patterns of lineage programs in single cells from fetal liver LSK subpopulations. Cells were scored as expressing MkE, GM, and/or lymphoid (Ly) programs based on the expression of one or more lineage associated genes: MkE: Gata1, VWF, and Epor; GM: Csf3r and Mpo; lymphoid: Rag1, sterile IgH transcript, and Il7r. Mean plus or minus SEM values from 2 experiments, with 88 single cells of each cell population investigated in each experiment, except for LSKFlt3− cells in which 88 cells were evaluated in total.

Combined GM and lymphoid transcriptional priming in LSKFlt3hiMpl BM cells. (A) Coexpression patterns of transcriptional lineage programs in single cells from BM LSK subpopulations. Cells were scored as expressing GM and/or lymphoid (Ly) programs based on the expression of one or more lineage-associated genes: GM: Csf3r and Mpo; lymphoid: Rag1, sterile IgH transcript, and Il7r. Mean plus or minus SEM values from 2 experiments with 88 cells investigated in each experiment. (B) Coexpression pattern of Mpl and Flt3 on Kit-enriched, lineage-negative (Lin), Sca-1+ and Kit+ (LSK) fetal liver (day E14.5-E15.5) cells. Gates denote the sorting strategies used to purify LSKFlt3hiMpl (Flt3hiMpl), LSKFlt3hiMplhi (Flt3hiMplhi), and LSKFlt3 cells. Percentages indicate mean quadrant frequencies within LSK cells from 4 experiments. Panels below show typical purity analysis for LSKFlt3hiMpl and LSKFlt3hiMplhi cells. (C) In vitro Mk potential of fetal liver LSKFlt3, LSKFlt3hiMplhi, and LSKFlt3hiMpl cells, investigated as described in “Methods” after 8 days of culture. Mean plus or minus SEM values from 4 experiments. (D) Coexpression patterns of lineage programs in single cells from fetal liver LSK subpopulations. Cells were scored as expressing MkE, GM, and/or lymphoid (Ly) programs based on the expression of one or more lineage associated genes: MkE: Gata1, VWF, and Epor; GM: Csf3r and Mpo; lymphoid: Rag1, sterile IgH transcript, and Il7r. Mean plus or minus SEM values from 2 experiments, with 88 single cells of each cell population investigated in each experiment, except for LSKFlt3 cells in which 88 cells were evaluated in total.

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