Anti cd9

The colon mucosa and splenic mononuclear cells were homogenized in modified RIPA lysis buffer supplemented with proteinase inhibitors (Vazyme biotech, Shanghai, China). Fifty micrograms of protein samples was separated on a 10% SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After electrophoresis, the proteins were transferred to PVDF (polyvinylidene difluoride) membrane (Millipore, IPVH00010, USA). The membrane was blocked in 5% nonfat milk for 1 h at room temperature and incubated with the corresponding primary antibodies at 4°C overnight. The sources of the antibodies were anti-IL-7 (Santa Cruz Biotechnology, USA), anti-CD9 (Bioworld Technology, USA), anti-CD63 (Bioworld Technology, USA), anti-CD81 (Epitomics, USA), and anti-β-actin (Santa Cruz Biotechnology). The next day, the blots were incubated with the secondary antibodies for 1 h at room temperature. The blots were visualized by chemiluminescence (Millipore, USA) and detected by using the imaging software (GE Healthcare, Life Sciences, USA).

RAW264.7 cells were obtained from the American Type Culture Collection (Rockville, MD, USA). WKYMVm and WRWWWW (WRW4) with purity > 95% was synthesized by GL Biochem and was dissolved in acetonitrile to prepare stock concentrations of 0.1, 1, 2, 5 and 10 μmol/L. Dulbecco's Modified Eagle's Medium (DMEM), alpha minimum essential medium (α‐MEM) and foetal bovine serum (FBS) were purchased from Invitrogen‐Gibco. RANKL and soluble mouse recombinant M‐CSF were obtained from R&D Systems. The Acid Phosphatase, Leukocyte (TRAP) assay kit and Actin Cytoskeleton and Focal Adhesion (FAK) Staining Kit were purchased from Sigma (Sigma‐Aldrich) and Merck, respectively. The Cell Counting Kit‐8 (CCK‐8) was purchased from Beyotime Biotechnology. Anti‐phospho‐NF‐kBp65, anti‐NF‐kBp65, anti‐STAT3, anti‐phospho‐STAT3 (Ser727), anti‐CTSK, anti‐c‐Fos, anti‐NFATc1, anti‐CD9, anti‐gp130, anti‐IL6 and anti‐ß‐actin antibodies were purchased from Bioworld Technology. Mouse IL‐1β, IL‐6 and TNF‐α ELISA kits were procured from Novus.

Angelicin with purity > 95% was purchased from Med Chem Express (Monmouth Junction, NJ, United States) and dissolved in DMSO. Alpha Minimum Eagle’s Medium (alpha-MEM), penicillin (10,000 units/ml), streptomycin (10,000 mg/ml) and fetal bovine serum (FBS) were purchased from Hyclone (Waltham, MA, United States). Soluble mouse recombinant M-CSF was purchased obtained from R&D system (Minneapolis, MN, United States). Interleukin (IL)-4, IL-13, LPS, and IFN-γ were purchased from PeproTech Inc. (Rocky Hill, NJ, United States). The Cell Counting Kit-8 (CCK-8) was purchased from Med Chem Express (Monmouth Junction, NJ, United States). Anti-CD9, anti-gp130, anti-STAT3, anti-phospho-STAT3 (Ser727), anti-CD206, anti-iNOS, anti-Arg-1, anti-β-actin antibodies were purchased from Bioworld Technology (St. Louis Park, MN, United States).