IL-10 modulates inflammasome-driven osteoclastogenesis ex vivo and during antigen-induced arthritis (AIA). (A) Correlation of Nlrp3 and Casp1 with Acp5 in mRNA derived from the synovium of IL-10KO mice at day 28 post AIA. Values are relative to 18S ribosomal RNA (n = 8-9). (B) Bone marrow cells from WT mice were cultured with MCSF in the presence (closed and open circles, left, black and grey bars, right) or absence (white bars, right) of RANKL and IL-10 (30 ng/ml) as indicated (open circles, left, grey bars, right). Values are derived from five independent fields of view from an experiment performed in triplicate. Representative pictures of cells treated with MCSF (left), MCSF and RANKL (middle) and MCSF, RANKL and IL-10 (right). Data was accumulated from two separate experiments. (Scale bar: 300 μm.) Expression of osteoclast markers; Acp5 and Ctsk, were determined by qPCR and compared against 18S ribosomal RNA. (C) Bone marrow cells from IL-10KO mice were cultured RANKL and MCSF. Osteoclast formation was determined in the presence of vehicle alone (closed circles), 30 μm of CRID3 (open circles, left) or 25 μg/ml glibenclamide (open circles, right). Data is representative of three experiments performed in triplicate (*P <0.05). Scale bar 300 μm. CRID3, cytokine release inhibitory drug-3; IL-10KO, interleukin-10-deficient; MCSF, macrophage colony-stimulating factor; qPCR, quantitative real-time PCR; RANKL, receptor activator of nuclear factor-κβ ligand; WT, wild-type.