Raw264.7 cells

BMDM and neutrophils were isolated from mouse bone marrow as previously described (14 (link)). BMDM were cultured with Dulbecco’s modified Eagle medium (Gibco, USA) supplemented with M-CSF at 10 ng/mL (PeproTech, Inc., USA). Neutrophils were purified using microbeads (Miltenyi Biotec, Germany). Peritoneal macrophages were collected through peritoneal lavage, as previously described (14 (link)). Murine RAW264.7 cells were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology (Shanghai, China).

Acteoside was isolated from C. deserticola by using the above method (Figure 8, UHPLC ≥ 98%); RAW264.7 cells were obtained from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. Shanghai, China; estradiol valerate was purchased from Delpharm Lille S.A.S, Paris, France; BGP, DPD, and TRAP crosslinks Elisa kits were bought from Xinyu Biological Engineering Co. Ltd., Shanghai, China; cathepsin K Elisa kit was purchased from Mountain View, CA, USA; MCSF and RANKL were provided by Pepro Tech Inc. American; total protein extraction kit and bicinchoninic acid (BCA) protein quantization kit were supplied by Ken Gen Biotech. Co. Ltd., Nanjing, China; primary antibodies including TRAF6, RANKL, RANK, NFKBIA, PI3K, AKT, IKKβ, NFAT2, c-Fos, β-actin, and secondary antibodies of horseradish peroxidase-conjugated goat anti-rabbit IgG were offered by ZSGB-BIO, Beijing, China; all other reagents were of analytical purity.

The murine primary BMMSCs were obtained from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (#ZQ0465, Shanghai, China). BMMSCs were cultured in DMEM/F12 (1:1) medium (#11320033, Gibco, US) containing 10% fetal bovine serum (FBS) and 1% double antibiotics (penicillin/streptomycin mix) (#10378016, Gibco, USA). The BMMSCs were maintained in a humidified atmosphere of 5% CO₂ at 37 °C. RAW264.7 cells were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (#ZQ0098, Shanghai, China), cultured with DMEM supplemented with 10% FBS and 1% antibiotics, and incubated in a humidified atmosphere of 5% CO₂ at 37 °C. For osteoclast differentiation induction, RAW264.7 cells were treated with 50 ng/mL RANKL (#CJ94, Novoprotein, Beijing, China).

RAW 264.7 cells and the corresponding complete medium were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China). Cells were cultured at 5% CO₂ and 37 °C in a humidified cell incubator machine. The experiment was divided into five groups: the PBS group (control), cells treated with PBS; the LPS group (LPS), cells treated with LPS (1 μg/ml) for 12 h; the LPS+MSC-Exo groups, cells treated with exosomes (10, 20, 40 μg/ml) for 24 h after the treatment of LPS.

RAW264.7 cells (Zhong Qiao Xin Zhou Biotechnology Co., Ltd., China, cat: ZQ0098) were cultured in the Dulbecco’s modified Eagle’s medium (DMEM) (Corning, cat: 10–013–CV) supplemented with 10% heat inactivated FBS (ExCell Bio, China, cat: FSS050) and 1% penicillin/streptomycin (Corning, cat: 30-002-Cl). THP-1 cells (Zhong Qiao Xin Zhou Biotechnology Co., Ltd, China, cat: ZQ0086) were cultured in RPMI-1640 medium (Corning, cat: 10-040-CV) supplemented with 10% heat inactivated FBS and 1% penicillin/streptomycin as mentioned above. THP-1 cells were differentiated (dTHP1) by incubation with 150 nM phorbol-12-myristate-13-acetate (PMA) (Sigma, cat: P8139) in complete medium for 24 h followed by 24 h PMA-free and serum-free medium treatment to reduce cell detachment (Spano et al., 2013 (link)).

The Clitocybe squamulosa were provided by the Edible Fungi Center of Shanxi Agricultural University (Shanxi Province, China). Mouse monocyte macrophage RAW264.7 cells were purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd (Shanghai Province, China). DEAE-cellulose-52 and Sephacryl S-400 were purchased from Solarbio (Beijing, China) and GE Healthcare (Chicago, IL, United States). All other chemical reagents were of analytical grade.