Figure 5

Abundance of Axin, Dvl2 and β-catenin in response to Wnt3a: effects of okadaic acid. F9 cells expressing Rfz1 were pretreated with or without 40 nM OA for 1 hr and then stimulated with Wnt3a in the presence or absence of OA for the indicated time periods. Cells were harvested and fractionated to subcellular fractions highly enriched in plasma membrane (PM), cytoplasm (CY), or nuclei (NU), as described in the Methods. Each fraction was separated by SDS-PAGE and the resolved protein transferred to blots stained with anti-Axin, anti-Dvl2 or anti-β-catenin antibodies. The enrichment of the subcellular fractions was established by staining immunoblots with antibodies to well known protein markers for PM, CY, and NU subcellular fractions. Left-handed panel displays the quantitative analysis of the blots for Wnt3a alone (blue line) or Wnt3a in the presence of OA (pink line). The data are displayed as the mean values "% of control" (i.e., t = 0 then set to 100%). Right-handed panel displays immunoblots. The results shown are derived from a single experiment, representative of two additional experiments.