Figure 3

Analysis of the Wnt signalling pathway in human primary osteoblasts following resistin stimulation. (A) Canonical Wnt signalling pathway gene expression following resistin stimulation. Primary human osteoblasts cells were stimulated with 500 ng/ml recombinant resistin for 24 hours prior to gene analysis, with 14 genes shown to be up-regulated (**p < 0.01, *p < 0.05). (B) IPA analysis of Wnt genes differentially expressed >1.4 fold change with resistin stimulation and the predicted increase in β-catenin activity and expression. Orange arrows = predicted activation, grey arrows = effect not predicted. (C) Representative images of nuclear translocation of β-catenin in human osteoblasts following i) media only ii) 30 min of resistin stimulation (500 ng/ml). Green = α-Actin, Blue = Hoechst, and Red = β-Catenin (n = 3). (D) Quantification of nuclear translocation of β-catenin in human primary osteoblasts using Image J software. Percentage nuclear β-catenin refers to the subset within the area of hoechst staining compared to the whole cell and is an average of 6 images from 3 OA patient cell lines (n = 3).