Figure 2.
Figure 2. Mixed myeloid composition of myeloproliferative disease in CD11cΔMHCII mice. WT and CD11cΔMHCII mice were assessed by flow cytometry at 12 weeks, 20 weeks, 30 weeks, and 1 year of age. (A) t-SNE projection of splenocytes of 1-year-old mice stained with myeloid markers (CD11b, Gr1, Ly6C, CD11c, and F4/80). Cells are grouped into clusters using FlowSOM, with blue representing WT splenocytes and red representing CD11cΔMHCII splenocytes. Marker composition of the 2 identified clusters (neutrophils and myeloid precursor-like cells) predominantly populated by CD11cΔMHCII splenocytes. (B) Absolute number of polymorphonuclear neutrophils (PMNs; defined as CD11b+Gr1high) in the spleen at 12 weeks (n = 11, 13), 20 weeks (n = 22, 16), 30 weeks (n = 18, 14), and 1 year (n = 22, 23) of age. The dotted line represents the threshold (2 standard deviations above the mean of WT mice) of neutropenia. Median and individual data points are shown. (C) Cumulative incidence plot for neutrophilia over time in WT and CD11cΔMHCII mice using Kaplan-Meier curve; all mice were censored at the time of analysis. The log-rank test was used for comparison between curves. (D) Absolute number of neutrophil-like precursor cells in the spleen (defined as CD11b+Gr1intLyC6intCD11c−) at 12 weeks (n = 9, 11), 20 weeks (n = 19, 16), 30 weeks (n = 12, 7), and 1 year (n = 22, 23) of age. The dotted line represents the threshold (2 standard deviations above the mean of WT mice) of increased myeloid precursor-like cells. Median and individual data points are shown. (E) Cumulative incidence plot for development of increased myeloid precursor-like cells over time in WT and CD11cΔMHCII mice using Kaplan-Meier curve; all mice were censored at the time of analysis. The log-rank test was used for comparison between curves. Absolute number (F) and percentage (G) of splenic hematopoietic stem cells (HSCs; defined as Lin−[TER119, CD3, NK1.1, CD11b, B220] Sca1+cKit+) in the spleen at 12 weeks (n = 9, 11), 20 weeks (n = 18, 14), 30 weeks (n = 8, 4), and 1 year (n = 20, 17) of age. Median and individual data points are shown.

Mixed myeloid composition of myeloproliferative disease in CD11cΔMHCIImice. WT and CD11cΔMHCII mice were assessed by flow cytometry at 12 weeks, 20 weeks, 30 weeks, and 1 year of age. (A) t-SNE projection of splenocytes of 1-year-old mice stained with myeloid markers (CD11b, Gr1, Ly6C, CD11c, and F4/80). Cells are grouped into clusters using FlowSOM, with blue representing WT splenocytes and red representing CD11cΔMHCII splenocytes. Marker composition of the 2 identified clusters (neutrophils and myeloid precursor-like cells) predominantly populated by CD11cΔMHCII splenocytes. (B) Absolute number of polymorphonuclear neutrophils (PMNs; defined as CD11b+Gr1high) in the spleen at 12 weeks (n = 11, 13), 20 weeks (n = 22, 16), 30 weeks (n = 18, 14), and 1 year (n = 22, 23) of age. The dotted line represents the threshold (2 standard deviations above the mean of WT mice) of neutropenia. Median and individual data points are shown. (C) Cumulative incidence plot for neutrophilia over time in WT and CD11cΔMHCII mice using Kaplan-Meier curve; all mice were censored at the time of analysis. The log-rank test was used for comparison between curves. (D) Absolute number of neutrophil-like precursor cells in the spleen (defined as CD11b+Gr1intLyC6intCD11c−) at 12 weeks (n = 9, 11), 20 weeks (n = 19, 16), 30 weeks (n = 12, 7), and 1 year (n = 22, 23) of age. The dotted line represents the threshold (2 standard deviations above the mean of WT mice) of increased myeloid precursor-like cells. Median and individual data points are shown. (E) Cumulative incidence plot for development of increased myeloid precursor-like cells over time in WT and CD11cΔMHCII mice using Kaplan-Meier curve; all mice were censored at the time of analysis. The log-rank test was used for comparison between curves. Absolute number (F) and percentage (G) of splenic hematopoietic stem cells (HSCs; defined as Lin−[TER119, CD3, NK1.1, CD11b, B220] Sca1+cKit+) in the spleen at 12 weeks (n = 9, 11), 20 weeks (n = 18, 14), 30 weeks (n = 8, 4), and 1 year (n = 20, 17) of age. Median and individual data points are shown.

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