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.