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Fig. 7. Molecular analysis of embryonic Carp-Lbh transgenic hearts. (A-H)
In situ hybridization showing downregulation of Anf mRNA (white
arrow) in Carp-Lbh transgenic hearts. Serial sections of E9.5
wild-type (wt; A,C,E,G) and transgenic (Tg; B,D,F,H) hearts were hybridized
with radioactive probes for Nkx2.5 (A,B), Tbx5 (C,D),
Gata4 (E,F) and Anf (G,H). (I,J) Lbh inhibits activation of
Anf-human growth hormone (Anf-hGH) reporter by cardiac transcription factors
in NIH 3T3 cells. (I) Dose-dependent suppression of Anf activation by Nkx2.5,
Tbx5 and Gata4 with increasing amounts of Lbh expression plasmid.
Values are expressed as percentage of total Anf-hGH reporter transactivation
by Nkx2.5, Tbx5 or Gata4 respectively. *P<0.001 (n=3)
versus Nkx2.5; **P<0.001 (n=3) versus Tbx5;
***P<0.05 (n=3) versus Gata4. (J) Inhibitory effect of
Lbh on synergistic transactivation of Anf by Nkx2.5 and Tbx5.
*P=0.008 (n=4) versus Nkx2.5 + Tbx5.
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