Fas ligand

To assess the contributions made by humoral factors, 5-wk-old CAG-eGFP mice were treated with TPO (Sigma-Aldrich) or IL-1α (R&D Systems or BioLegend) at dosages of 10 µg/mouse subcutaneously (SC) daily for 5 d or 70 µg/mouse SC daily for 3 d. The mice were visualized and examined at 6-wk-old, 7 d after the first treatment. Some mice were also treated with neutralizing antibodies against IL-1α (R&D Systems or BioLegend), IL-1R (BioLegend), isotype-matched control antibody (BioLegend), or Fas Ligand (5 µg/mouse i.v.; R&D Systems). All antibodies were administered at 100 µg/mouse i.p. daily for 3 d.
To assess the effect of acute inflammation, thioglycolate (Sigma-Aldrich) was administered i.p. (3 ml of 3% solution/mouse once). To examine the effect of caspase inhibition, mice were treated with the pan-caspase inhibitor Z-VAD (OMe)-FMK (3 mg/kg IP daily for 5 d; Merck Millipore). Clophosome (200 µl/mouse; FormuMax Scientific Inc.) was administered i.p. to deplete macrophages.
To obtain chimeric mice, 6-wk-old WT mice were lethally irradiated at a dose of 9.5 Gy, after which they were transplanted with BM total cells from age-matched IL-1R^−/−/CAG-eGFP, IL-1α^−/−/CAG-eGFP, or control (IL-1R^+/+/CAG-eGFP or IL-1α^+/+/CAG-eGFP) mice. 6 wk after transplantation, BM dynamics were scored.