Fig 2: Gas6 blockade does not affect the composition or activation status of myeloid cells and T cells in pancreatic tumors. (A) Mass cytometry quantification of CD11b + myeloid cells, Ly6C high/Ly6C low monocytes/MDSCs, Ly6G high/Ly6C low neutrophils/MDSCs, F4/80+ macrophages, MHCII+ macrophages, CD206+ macrophages, and PD-L1+ macrophages in mouse pancreatic tumors treated with control IgG (n = 3) or anti-Gas6 neutralizing antibody (n = 4). Values shown are mean and SEM. n.s. no statistically significant differences, using unpaired student T-test. (B) Mass cytometry quantification of CD3+ T cells, CD4+ T cells, CD4+/CD25+ regulatory T cells (Tregs), CD8+ T cells, CD69+/CD8+ T cells and PD-1+/CD8+ T cells in mouse pancreatic tumors treated with control IgG (n = 3) or anti-Gas6 neutralizing antibody (n = 4). Values shown are mean and SEM. n.s. no statistically significant differences, using unpaired student T-test. Graphs were generated with ViSNE data using Cytobank software.

Fig 3: MDSCs constantly accumulated in the liver during CRC process.A Diagrams of immune cells by flow cytometry sorting. CD45+ cells were first sorted and were stained with CD3 and CD4 antibodies; CD8+ T cells were stained with CD3 and CD8 antibodies; dendritic cells were stained with CD11b and CD11c antibodies; macrophages were stained with F4/80 and CD11b antibodies; natural killer cells were stained with NK1.1 antibodies; MDSCs were stained with CD11b, Gr-1 or Ly6G or Ly6C antibodies. B Analysis of the numbers of immune cells in CRC liver metastases. C Analysis of the numbers of Mo-MDSCs or PMN-MDSCs in CRC liver metastases. **P < 0.01, ***P < 0.001. D–F Analyses of the MDSCs percent in CRC patients’ blood. Normal (n = 20), CRC with metastasis (n = 30), CRC without metastasis (n = 30). Error bars represent mean ± SD from at least three independent experiments. **P < 0.01, ***P < 0.001.

Fig 4: The proportions of leukocytes other than neutrophils were not increased after anesthesia/surgery. Representative flow cytometry data of blood showed the percentage of macrophages (CD45+ CD11b+ F4/80+), B cells (CD45+ CD19+), T cells (CD45+ CD3+), monocytes (CD45+ CD11b+ F4/80– Ly6G– Ly6C+), NK cells (CD45+ NK1.1+) and dendritic cells (CD45+ CD11c+) at 24 h after anesthesia/surgery. The percentage of NK cells (E, P = 0.0396) were relatively decreased because of the increase of neutrophils after anesthesia/surgery compared with control group, while no significant difference was found in the percentage of macrophages (A, P = 0.8749), B cells (B, P = 0.0609), T cells (C, P = 0.1089), monocytes (D, P = 0.2883), and dendritic cells (F, P = 0.0854). (n = 3–4 per group). Data are expressed as means ± SE. *P < 0.05 compared with that of the control group.

Fig 5: House dust mite exposure induces airway eosinophilic and T helper 2-associated in?ammation. (A) Flow cytometry analysis of eosinophils in the lung. (B) Changes of CD45+CD11b+SiglecF+Ly6C in eosinophil proportions are presented using a bar graph. Relative expression levels of (C) IgE, (D) IL-5 and (E) IL-13 in BALF were assessed. *P<0.05, **P<0.01 and ***P<0.001 compared with the control group. BALF, bronchoalveolar lavage fluid.