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(A) Bone marrow-derived macrophages (BMM) from WT and Fireko mice were generated by culture in <t>CSF2</t> for 7 days followed by staining and flow cytometry analysis. F4/80/CD11b + cells were gated based on MHCII expression. The relative proportions of MHCII High and Low subsets and their CSF1R MFI are shown. Data show mean and standard deviation, 2-way ANOVA with Sidak’s multiple comparison test. (B) CSF2-derived BMM were incubated with pH Rodo E.coli bioparticles (BP) for 1 h followed by surface staining and analysis by flow cytometry for BP uptake (%) and median fluorescence intensity (MFI) in cell subsets. Data show mean and standard deviation. (C) Representative images of bone marrow from WT or Fireko mice cultured in CSF1, CSF2 or both for 3 days prior to the addition of RANKL and continued culture until day 7. Cells were stained for tartrate-resistant acid phosphatase (TRAP, purple)
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(A) Bone marrow-derived macrophages (BMM) from WT and Fireko mice were generated by culture in CSF2 for 7 days followed by staining and flow cytometry analysis. F4/80/CD11b + cells were gated based on MHCII expression. The relative proportions of MHCII High and Low subsets and their CSF1R MFI are shown. Data show mean and standard deviation, 2-way ANOVA with Sidak’s multiple comparison test. (B) CSF2-derived BMM were incubated with pH Rodo E.coli bioparticles (BP) for 1 h followed by surface staining and analysis by flow cytometry for BP uptake (%) and median fluorescence intensity (MFI) in cell subsets. Data show mean and standard deviation. (C) Representative images of bone marrow from WT or Fireko mice cultured in CSF1, CSF2 or both for 3 days prior to the addition of RANKL and continued culture until day 7. Cells were stained for tartrate-resistant acid phosphatase (TRAP, purple)

Journal: bioRxiv

Article Title: Depletion and replacement of tissue resident macrophages in mice with germ-line deletion of a conserved enhancer in the Csf1r locus

doi: 10.64898/2026.03.22.713539

Figure Lengend Snippet: (A) Bone marrow-derived macrophages (BMM) from WT and Fireko mice were generated by culture in CSF2 for 7 days followed by staining and flow cytometry analysis. F4/80/CD11b + cells were gated based on MHCII expression. The relative proportions of MHCII High and Low subsets and their CSF1R MFI are shown. Data show mean and standard deviation, 2-way ANOVA with Sidak’s multiple comparison test. (B) CSF2-derived BMM were incubated with pH Rodo E.coli bioparticles (BP) for 1 h followed by surface staining and analysis by flow cytometry for BP uptake (%) and median fluorescence intensity (MFI) in cell subsets. Data show mean and standard deviation. (C) Representative images of bone marrow from WT or Fireko mice cultured in CSF1, CSF2 or both for 3 days prior to the addition of RANKL and continued culture until day 7. Cells were stained for tartrate-resistant acid phosphatase (TRAP, purple)

Article Snippet: 10 7 BM cells were seeded on 100 mm square bacteriological plastic (Sterilin, Thermo-Fisher, Australia) in 25 ml of complete medium (RPMI + 10% FBS, 25 U/mL penicillin, and 25 μg/mL streptomycin (Gibco, Thermo-Fisher, Australia) and differentiated for 7 days with the addition of recombinant CSF1-Fc (100 ng/ml) or recombinant mouse CSF2 (GM-CSF, R&D Systems, Minneapolis, Mn, USA; 50 ng/ml).

Techniques: Derivative Assay, Generated, Staining, Flow Cytometry, Expressing, Standard Deviation, Comparison, Incubation, Fluorescence, Cell Culture