Five-week-old male C57BL6/J mice were obtained from Japan CLEA (Japan). Bone marrow cells were collected from the tibiae and femur, and were cultured in M-CSF (50 ng/ml) for 3 days in culture dishes containing DMEM (Wako) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, United States). After 3 days, floating cells were removed by rinsing with phosphate-buffered saline (PBS), and the attached cells were used as mBMMs (Horibe et al., 2013; Tevlin et al., 2014). Attached mBMMs were sprayed with PBS to remove the culture dish mechanically to use for the following in vitro experiments. This animal experiment was performed in accordance with the Guidelines for Proper Conduct of Animal Experiments of the Science Councils of Japan and approved by the Animal Research Control Committee of Okayama University (approval no. OKU-2016277).

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