Additional file 2: of Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state

Figure S2. Indirect effects of MSU crystal-stimulated RAW264.7 macrophage conditioned medium on MLO-Y4 cell viability. RAW264.7 macrophages were cultured with or without 0.5 mg/mL MSU crystals for 24 h for preparation of MSU crystal-stimulated conditioned medium and control conditioned medium, respectively. Conditioned medium preparations were added to MLO-Y4 cells at different concentrations (5%, 20%, and 40% final concentration in a well) for 24 h. The alamarBlue® assay was used to determine MLO-Y4 cell viability 24 h and 48 h after the addition of conditioned medium. Data shown are pooled from three biological repeats and are presented as mean (SEM), two-way ANOVA: PInteraction = 0.16; PTime = 0.74; and PConditioned media concentration = 0.17. (JPG 152 kb