Figure 4: Succinate stimulates osteoclastogenesis via SUCNR1.
From: Succinate and its G-protein-coupled receptor stimulates osteoclastogenesis
As described in the method, ficoll-processed non-adherent bone marrow cells were seeded at 20,000 per well in 96-well plate and stimulated with the indicated concentration of cytokines (M-CSF, 30 ng ml−1 from day 1; RANKL, 50 ng ml−1 from day 3); fresh cytokines were added every other day. (a) Western blotting of SUCNR1 in various cell lineages from bone. Representative TRAP staining images and analysis of (b,c) WT mice bone marrow in vitro OC cultures treated with 1 mM succinate or control in the presence of SUCNR1 antagonist 4c at indicated concentrations. Scale bar, 200 μm. (d,e) Succinate effects in the OC cultures derived from bone marrow of WT and SUCNR1 KO mice, scale bar 250 μm. Data show mean±s.e.m. of triplicates. **P<0.01, ***P<0.001 according to Bonferroni post hoc test after ANOVA, n=5.
