bves regulates the development of the outflow tract via SHF. (A) SHF gene expression was detected in bves morphants by qRT-PCR at 48 hpf. (B) Overexpression of BVES led to upregulation of transcriptional activity at the promoters of GATA4, NKX2.5 and HAND2, downregulation of transcriptional activity at the promoters of TBX1, and no change in transcriptional activity at the promoters of TBX20 and MEF2C. pCMV-Vector, empty vector of pCMV-Myc; pCMV-BVES, the vector overexpressing BVES; pGL3-Vector, empty vector of pGL3-Bias; pGL3-Promotor, vector containing the promotor of gene. Red *: significance analysis with pGL3-Vector group; Black *: significance analysis with pGL3-Promotor + pCMV-Vector group. *p < 0.05; **p < 0.01; ***p < 0.001. (C) Outflow tract phenotype of the wild type, bves Mo injected, bves Mo and nkx2.5 mRNA coinjected in end-systolic at 72 hpf in Tg (flia:eGFP). The words that marked by red color is the width of outflow tract. ca, caudal; cr, cranial; r, right; l, left. Scale bar = 15 μm. (D) The width of the outflow tract at the end-systolic stage in (C). Compared with the WT and bves Mo, the width of the outflow tract was partially rescued by nkx2.5 mRNA (WT, 9.5 μm, bves Mo, 5.0 μm, p < 0.01 compared with WT; bves Mo + nkx2.5 mRNA, 6.3 μm, p < 0.01 compared with WT, p < 0.05 compared with bves Mo). n, number of samples. OT, outflow tract. *Significance analysis with WT; #Significance analysis with bves Mo. #p < 0.01; **p < 0.01. The error bar shows the mean and SD. WT: wild type; bves Mo: bves morphants; bves Mo + nkx2.5 mRNA, coinject bves morpholino and nkx2.5 mRNA.
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