Additional file 2: Figure S1. of Generation mechanism of RANKL+ effector memory B cells: relevance to the pathogenesis of rheumatoid arthritis

Analysis of osteoclast differentiation using RAW-Venus1. (A) Differentiation into Venus positive pre-osteoclasts and osteoclasts were induced by RANKL in a dose-dependent manner. TNF-α exerted synergistic effects on RANKL-induced osteoclast differentiation (magnification × 20). (B) Both anti-RANKL Ab and OPG strongly reduced the number of Venus positive cells, indicated the inhibition of osteoclast differentiation (magnification × 20). (C) Venus positive cells were tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (magnification × 40). Cells were stained using a commercial TRAP assay (Sigma-Aldrich, St Louis, MO, USA). (A-C) Cells were cultured at 4.5 × 104 cells/ml for 3 days in a 96-well plate. No stim No stimulation. (PDF 304 kb)