Fig. 2.
Biglycan-deficient calvarial cells retain less Wnt3a in cell layers and demonstrate both reduced Wnt-induced phosphorylation of LRP6 and reduced β-catenin/TCF-mediated transcriptional activity. (A) Immunoblot analysis for biglycan core protein after treatment of an equal volume of lysate proteins obtained from WT calvarial cell layers with chondroitinase ABC. (B) Immunoblot analysis for Wnt3a and phosphorylation of LRP6 (pLRP6S1490) on protein extracts obtained from WT and biglycan-deficient calvarial cells after treatment with Wnt3a. (C) Relative expression levels of Wnt target genes measured by real-time RT-PCR in WT and biglycan-deficient calvarial cells after treatment with Wnt3a. Values represent mean relative expression ± SD (n = 3), shown as Wnt3a-induced fold change in gene expression. Gene expression was normalized to untreated controls at each time point (indicated by the dotted line). *P < 0.05; **P < 0.01; ***P < 0.001.