Fig. 5

CB2 antagonizes TLR-4 signaling in macrophages. a The PMs were double-stained with TLR4 (Red) and CB2 (Green) antibodies by immunofluorescence. The nucleus was visualized by DAPI staining (Blue). This test was repeated three times. Representative images were shown. Scale bars, 40 μm. b Protein–protein Interaction between CB2 and TLR4 in the mouse peritoneal macrophages treated with PBS or LPS (500 ng ml⁻¹) for 30 min. The proteins were immunoprecipitated with TLR4 antibody or IgG as control. This test was repeated three times. Representative results were displayed. c LPS or AM630 inhibits, while 2-AG induces the Interaction between CB2 and TLR4 in macrophages. The mouse PMs described in f were harvested and the proteins were immunoprecipitated with CB2 antibody. d Relative mRNA levels of inflammatory cytokines in the TAMs from the WT, Tg^CB2, TLR4^KO or Tg^CB2 + TLR4^KO mice bearing MC-38 tumors for 3 weeks. (n = 5) Data in j are means ± s.e.ms. (*P < 0.05, **P < 0.01, ***P < 0.005; student’s t-test; ns not significant)