Fig. 5

Tandem CSF1-Fc and G-CSF treatment had modest cumulative impacts on myeloid cells. a Schematic of tandem CSF1-Fc plus G-CSF treatment regimen administered to C57BL/6 non-transgenic mice. Briefly, mice were divided into 4 treatment groups: (1) once daily saline for 4 days followed by bi-daily saline treatment initiated 14 days later (saline + saline); (2) once daily CSF1-Fc for 4 days followed by bi-daily saline (CSF1-Fc + saline); (3) once daily saline for 4 days followed by bi-daily G-CSF treatment (saline + G-CSF); and (4) once daily CSF1-Fc for 4 days followed by bi-daily G-CSF treatment (CSF1-Fc + G-CSF). Tissues were harvested 17 days post-first CSF1-Fc injection. b Representative immunohistochemistry anti-F4/80 staining (brown) in femoral sections of mice treated as described above. F4/80⁺ BM resident macrophages (brown) are lining the endosteal (arrowheads) and perivascular (arrow) regions of the BM. Sections were counterstained with hematoxylin (blue) and taken at 600X magnification; scale bar = 20 µm. c Quantification of percent area of F4/80 staining in the femur of mice treated as above. d Representative immunohistochemistry anti-F4/80 staining (brown) in splenic sections of mice treated as above. Section were counterstained with hematoxylin (blue) and taken at 100X magnification; scale bar = 500 µm. Inset at 600X magnification; scale bar = 20 µm. e Quantification of percent area of F4/80 staining in the spleen of mice treated as above. f Weights of spleen of mice treated as above. g–j Flow cytometry analysis of the percentage of F4/80⁺Ly6G^negVCAM⁺CD115⁺CD11b⁺ monocytes (Mo) in BM (g) and spleen (h) or CD11b⁺Ly6G⁺ granulocytes (Grans) in BM (i) and spleen (j) of mice treated with either saline + G-CSF or CSF1-Fc + G-CSF. Each data point represents a separate mouse, and bars are mean ± SD. Statistical analysis was performed using one-way ANOVA Tukey’s multiple comparison test and unpaired Student's t test where ****p < 0.0001, ***p < 0.001, *p < 0.05