Male C57BL/6 (Charles River, Germany) and bone marrow chimera mice (see below) weighing 25.4 ± 3.8 g were maintained under standardized conditions (12:12 h light-dark cycle, 40–70% relative humidity, and constant ambient temperature of 22 ± 2 °C), with free access to standard rodent chow (Akronom Kft., Budapest, Hungary) and tap water. The experiments were approved by the Animal Ethics Committee of Semmelweis University and the Pest County Government Office (PE/EA/2202-5/2017). All procedures were performed in accordance with the Hungarian Law on the protection and welfare of animals and the EU Directive 2010/63/EU for animal experiments.
For testing the role of neutrophil granulocytes, bone marrow chimeras with a neutrophil-deficient Mcl1flox/floxLysMCre/Cre hematopoietic system were used [47 (link),48 (link)]. Briefly, C57BL/6 recipients were irradiated with 11.5 Gy gamma rays using a 137Cs source and injected intravenously with unfractionated bone marrow cells from wild-type (WT) or Mcl1flox/floxLysMCre/Cre (referred to as Mcl-1ΔMyelo) mice on C57BL/6 background. The number of circulating neutrophils was counted by flow cytometry four weeks later. Neutrophils were defined as Ly6G-positive cells within a typical forward and side-scatter gate.
For testing the role of neutrophil granulocytes, bone marrow chimeras with a neutrophil-deficient Mcl1flox/floxLysMCre/Cre hematopoietic system were used [47 (link),48 (link)]. Briefly, C57BL/6 recipients were irradiated with 11.5 Gy gamma rays using a 137Cs source and injected intravenously with unfractionated bone marrow cells from wild-type (WT) or Mcl1flox/floxLysMCre/Cre (referred to as Mcl-1ΔMyelo) mice on C57BL/6 background. The number of circulating neutrophils was counted by flow cytometry four weeks later. Neutrophils were defined as Ly6G-positive cells within a typical forward and side-scatter gate.
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