Dvl-3 as a marker of Wnt pathway activation. Line 30F (left panel) or co-cultures of Lines 31E + 30F (right panel) stimulated with ectopic Wnt-3a respond with hyperphosphorylation of Dvl-3. Following treatment with Wnt-3a for the indicated times, cells were lysed with Triton X-100 Lysis Buffer and cell extracts were subjected to Western blot analysis. Hyperphosphorylation of Dvl-3 is evident by a consistent relative increase in the intensity of the upper band of the doublet. Alkaline phosphatase treatment caused the loss of the upper band (data not shown).