Fig. 5. Effect of ß-eng expression on HC11 cell proliferation. Western blot analysis of three replicate HC11 cell cultures infected with ß-catenin, ß-eng or mock-infected control (A). ß-catenin and ß-eng transgene expression in infected cells detected by an antibody against the mycepitope tag. Anti-ß-catenin antibody detected endogenous ß-catenin and served as a loading control. Anti-cyclin-D1 antibody detected no change in the level of cyclin D1 protein, whereas activated MAPK was upregulated compared with the total MAPK protein level in response to ß-eng expression. Quantitation of pMAPK activation by densitometry (C) revealed a 15-fold increase in activated MAPK compared with total MAPK. Proliferation in HC11 cells was measured by immunfluorescent detection of FITC-labelled BrdU incorporation. Quantitation of BrdU-labelled cells (B) revealed no change in proliferation in cells infected with ß-catenin or ß-eng compared with mock-infected cells.

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