Figure 2. Wnt3a stimulates fibroblasts proliferation, migration and gel contraction.

Confluent foreskin fibroblasts were infected with indicated expression vectors (Ad-GFP, Ad-Cat^ca or Ad-Wnt3a, 30 MOI) or transiently transfected with Fzd2, or siRNA specific for Wif1 or DKK2 or corresponding scrambled control siRNAs, followed by incubation for 24-48 h in media with Wnt3a (100 ng/ml) or LiCl (30 mM). A, B. RNA was isolated and examined by real-time qPCR. Results, normalized with Gapdh, are shown as means ± SD of triplicates from three independent experiments. * p < 0.05. C. Left panels, whole cell lysates were subjected to GST-ICAT pull-down assays (see Material and Methods). Active β-catenin binding to ICAT was probed with a pan-catenin antibody. Representative images. Right panels, fibroblasts were incubated with Wnt3a for 4 h, fixed and probed with DAPI (blue) and β-catenin antibody (green), and examined by immunofluorescence microscopy. Representative images, original magnification × 400. D. Fibroblast migration was monitored by measuring scratch width at three different sites in each sample. Results are shown as means ± SD of triplicates from an experiment representative of three. * p < 0.05. E. Proliferation assays. * p< 0.05. F. Gel contraction assays. Results expressed as the percentage of gel area compared to time 0 are shown as mean ± SD of a triplicates from representative experiment of three independent experiments. * p <0.05.