Fig. 3. (A) Sequence alignment of the EGF-CFC proteins: mouse Cripto (mCripto), human Cripto (hCripto), zebrafish Oep (Oep), human Cryptic (hCryptic), mouse Cryptic (mCryptic), frog FRL1 (FRL1) and chicken Cripto (cCripto). The positions of the missense mutations (red residues) are shown above the alignment. Red and blue lines indicate amino acids deleted in cripto-His and in the EGF-CFC-His constructs, respectively. Blue and green shaded areas indicate the EGF and the CFC domains, respectively. Conserved residues in the EGF-CFC domain are indicated by asterisks. The white arrow indicates the predicted cleavage site of signal peptide in mouse Cripto. Black arrows indicate processing sites of recombinant Cripto-His expressed in 293T cells. (B) Western blot analysis of total lysates from 293T cells transfected with Cripto wild-type and mutant derivatives. Cells were cotransfected with Jun-Ha expression vector as internal control. The molecular mass of protein standards is indicated (kDa).
