|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 4. The response of Grsf1 and Fragilis2 to Wnt signaling and
shRNA silencing. (A) Co-culture of ES cells on lacZ and
Wnt1-expressing fibroblasts induces Grsf1 endogenous mRNA
expression in Wnt1-treated ES cells, but not Fragilis2 expression.
Gene-specific expression was normalized to Gapdh and the fold change was
calculated. Results of three independent co-culture experiments are shown.
Whole-mount in situ hybridization of wild-type and ß-catenin mutants
(B,C), and of wild-type and shRNA-silenced embryos (E,F) with Grsf1
and Fragilis2 probes at late-gastrulation stage. All embryos are
depicted in a lateral view, anterior to the left. Grsf1 and
Fragilis2 are strongly downregulated in the primitive streak of
ß-catenin mutant embryos (B,C) and knock-down embryos (E,F). Arrow in C
indicates remaining Fragilis2 expression at the base of the
allantois. (D) Northern blot analysis for Grsf1 and
Fragilis2 for eight different shRNA ES-cell lines (1-8) and wild-type
ES cells. 28S, 18S and 5.8S ribosomal RNAs are indicated as molecular length
markers, and the ethidium bromide (EtBr)-stained agarose gels served as
loading control.
|
