Figure 2 – G-CSF and immunosuppressive neutrophilic cells accumulate in the periphery of mice with progressing MOC1 tumors.

Representative flow cytometry dot plots of (A) bone marrow, (B) peripheral blood and (C) spleen samples (n=5 mice per timepoint) from non-tumor bearing mice (top dot plots) and day 38 tumor bearing mice (bottom dot plots) and connected line dot plots show the quantification of Ly6G+ or Ly6C+ myeloid cells as a percentage of live total leukocytes in tumor bearing mice during tumor progression. The concentration of cells in mice without tumors is shown as a dotted line in each plot. ***, P<0.001; significance was determined using multiple unpaired t-tests with each timepoint considered independently.

D, connected-line dot plots show the concentration of plasma growth factors in tumor bearing mice (n=5 mice per timepoint) during tumor progression. The plasma concentration of each factor in mice without tumors is shown as a dotted line in each plot.

E, representative flow cytometry dot plots (left) and dot plots showing quantification (right) of the percentage of CD31⁺CD45⁻ endothelial cells, CD31⁻CD45⁻ tumor cells, T cells, Ly6C⁺F4/80⁻ monocytes, Ly6C⁺F4/80⁺ macrophages, and Ly6G⁺ neutrophilic cells from the spleen or tumor of day 35 tumor-bearing mice (n=5) positive for G-CSF production.

F, representative CFSE-labelled T cell proliferation histograms and dot plots showing the quantification of the ability of Ly6G+ cells from the spleens or tumors of day 38 tumor bearing mice (TBM) or splenic Ly6G+ cells from non-tumor bearing mice or naïve total splenocytes to suppress T cell proliferation or IFNγ production. Significance determined with ANOVA multiple comparisons test.