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Fig. 6. Stabilized ß-catenin is insufficient for induction of
primitive streak-associated gene expression. (A) A2lox.sßcat
ES cells transiently transfected with the SUPER8xTOPFlash (TOP) or
SUPER8xFOPFlash (FOP) reporters were left untreated (no treatment), or
treated with either 1 µg/ml doxycycline (DOX) or LiCl for 18 hours. Whole
cell lysates were prepared and luciferase activity measured. Shown is the
luciferase activity normalized to a co-transfected Renilla luciferase
construct driven by the CMV promoter as previously described
(Ranganath et al., 1998).
(B) A2lox.sßcat ES cells were differentiated in SRM alone or in
the presence of Dkk1 (+Dkk1) and of the indicated concentration of doxycycline
(DOX). Dkk1 was added at the initiation of differentiation. Doxycycline was
added at day 1 of differentiation. Cells were harvested at day 3.5 of
differentiation and evaluated for expression of the indicated genes by RT-PCR.
Expression found in ES cells differentiated in SCM without doxycycline
represent positive controls.
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