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. 2019 Sep 5;49(11):2103–2110. doi: 10.1002/eji.201948124

Figure 1.

Figure 1

Flow cytometry analysis of NMR leukocytes. Cells were isolated from the NMR spleen, blood and bone marrow and stained as described in Materials and Methods and in Table 1. Cells were analysed by flow cytometry for: (A, B) distribution of size (forward scatter/FSC‐A) and granularity (side scatter/SSC‐A) by conventional flow cytometry. A total of five NMR samples were analysed in four independent experiments (A). Representative results of one of four experiments. (B) Data of four experiments with five NMR samples. (C) Expression of IgM and MHC‐II in G1‐G4 sub‐populations. (D) Size, granularity and expression of IgM, MHC‐II and CD14 analysed by imaging flow cytometry. Representative results of one of two experiments with a total of two NMR samples are shown. n.s. – non‐specific binding. Images also including cells from NMR blood and spleen are shown in Supporting Information Fig. S6. (E) Expression of CD3ε, CD8α and CXCR3 in G1 sub‐population. (F) Expression of CD14 and MHC‐II in G1‐G4 sub‐populations. (G) Expression of CD34 and c‐Kit/CD117 in G1‐G4 sub‐populations. (C, E‐G) Representative results of one of three experiments with a total of three NMR samples are shown. Additional samples from bone marrow (C, E, F) and blood (G) and corresponding unstained controls are shown in Supporting Information Figs. 5 (C), S8 (E), S9 (F) and S10 (G). (H) Percentages of granulocytes, CD3ε+ T cells, CD3ε+CD8α+ T cells, IgM+ B cells and CD14+MHC‐II+ monocytes/macrophages are shown. Data from three independent experiments are shown. N – number of analysed NMR samples. (B, H) Data are shown as mean ± standard deviation (SD). Student's t‐test was used for statistical analysis. The difference is considered statistically significant if p < 0.05. (I) Frequency of total CD3ε+ (upper panel) and cytotoxic CD3ε+CD8α+ (lower panel) T cells in NMR, mouse (Ms) and human (Hu) organs. Data from three independent experiments are shown for NMR (five and three samples were analysed accordingly for total CD3ε+ and cytotoxic CD3ε+CD8α+ T cells). Data for the mouse (averaged for CB17, C57BL/6 and BALB/c strains) and humans are taken from the literature sources19, 20, 21, 22. Data are shown as mean ± SD. Gating strategy is shown in Supporting Information Figs. 4 (C, E, F and G) and S6 (D).