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Fig. 3. TEAD4 is required for expression of some genes during preimplantation
development, but not others. (A) Embryos from
Tead4+/- heterozygous intercrosses were collected either
at E2.5 or E3.5 and immunostained with anti-CDX2 antibody (green). DNA was
labeled with DAPI (blue), and embryos were photographed under phase contrast
optics (Phase). Confocal images of DNA and CDX2 stains were merged. Although
the top panel appears to show an 8-cell embryo, there are four more nuclei
present outside the focal planes shown. Embryos were genotyped after imaging.
Wild-type and Tead4 heterozyotes were morphologically
indistinguishable, so only wild-type and Tead4 knockout embryos are
shown (total number of embryos examined: Tead4+/+,
Tead4+/-: 13 for E.2.5 and 14 for E3.5;
Tead4-/-: 3 at E2.5 and 4 at E3.5). (B) Embryos
from Tead4+/- heterozygous intercrosses were collected at
E3.5 and stained for OCT4 protein (green) as described in A. Typical examples
are shown of Tead4+/+ blastocysts (n=16), and of
Tead4-/- embryos (n=4). (C) Total RNA was
isolated from a single E3.5 blastocyst and a single E3.5 abnormal morula, and
RT-PCR was used to detect expression of the Tead4, Cdx2, Eomes, Fgfr2
and Oct4 genes. ß-actin RT-PCR products are shown as a
reference.
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