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Fig. 7. Expression of dominant negative Tead2 affects Pax3 expression. A Tead2
fusion protein including the Engrailed repressor domain was expressed in the
neural tube using the Wnt1 promoter. (A) Wild-type embryo with normal
morphology and Pax3 protein expression (green) in the dorsal neural tube (nt),
dorsal root ganglia (drg, dotted line) and somites. (B) Adjacent Haematoxylin
and Eosin-stained wild-type section. (C) Transgenic embryo with an abnormal
dorsal neural tube (nt) and small dorsal root ganglia (drg, dotted line) with
diminished Pax3 expression in neural tube and drg. Pax3 protein expression in
the somites is unaffected. (D) Adjacent Haematoxylin and Eosin stained
transgenic section. (E) Wild-type E10.5 embryo stained with anti-neurofilament
antibody (arrows) showing strong expression adjacent to neural tube (nt) and
dorsal root ganglia and weaker expression by enteric ganglia (eg) in the
hindgut (hg). (F) Neurofilament expression in transgenic embryo is weak in
regions adjacent to dorsal root ganglia (arrows) and absent from hindgut. (G)
Tead2-Engrailed inhibits Tead2 and YAP65-mediated activation of a luciferase
reporter construct containing NCE2 (NCE2-luciferase) in a dose-dependent
manner. Fold activation of luciferase activity is corrected for transfection
efficiency and performed as in Fig.
5. (H) Retinoic acid induces endogenous Pax3 protein expression
(red; detected by immunohistochemistry) in P19 cells transfected with a GFP
expression vector (green). (I) Co-transfection of a Tead2-Engrailed expression
vector with GFP inhibits Pax3 expression such that those cells expressing GFP
do not express Pax3. Nuclei are stained blue.
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