TABLE 1.
Plasmids used in this work
| Plasmid | Use | Reference |
|---|---|---|
| Cripto | ||
| pcDNA3-FLAG-Cripto | Mammalian expression of mouse Cripto (full-length coding region only) with FLAG epitope tag inserted at N terminus following signal sequence | This work |
| pcDNA3-HA-Cripto | Mammalian expression of mouse Cripto with HA epitope tag inserted at N terminus following signal sequence | This work |
| pFLAG-Cripto/IRES-GFP | Generation of stable clones expressing mouse Cripto (full-length coding region with FLAG epitope tag) | This work |
| pcDNA3-FLAG-Cripto(L75A/S77A/F78A) | Expression of triple point mutant in EGF motif of mouse Cripto | This work |
| pcDNA3-FLAG-Cripto(P82A/F85A/R88A) | Expression of triple point mutant in EGF motif of mouse Cripto | This work |
| pcDNA3-FLAG-Cripto(H104A/W107A/R116A) | Expression of triple point mutant in CFC motif of mouse Cripto | This work |
| pcDNA3-FLAG-Cripto(L122A/P126A/D134A) | Expression of triple point mutant in CFC motif of mouse Cripto | This work |
| pcDNA3-FLAG-Cripto(T72A) | Expression of O-fucosylation site mutant of mouse Cripto | This work |
| pcDNA3-HA-Cripto(T72A) | Expression of O-fucosylation site mutant of mouse Cripto | This work |
| Cryptic | ||
| pcDNA3-FLAG-Cryptic | Mammalian expression of mouse Cryptic (full-length coding region only) with FLAG epitope tag inserted at N terminus following signal sequence | This work |
| pEFneo-HA-hCryptic | Expression of human Cryptic with HA epitope tag inserted at N terminus following signal sequence | 3 |
| pEFneo-HA-hCryptic(R112C) | Expression of human Cryptic mutant associated with left-right laterality defects | 3 |
| pEFneo-HA-hCryptic(G174del1) | Expression of human Cryptic mutant associated with left-right laterality defects | 3 |
| pEFneo-HA-hCryptic(R78W) | Expression of human Cryptic mutant associated with left-right laterality defects | 3 |
| pEFneo-HA-hCryptic(R189C) | Expression of human Cryptic mutant associated with left-right laterality defects | 3 |
| oep | ||
| pcDNA3-FLAG-oep | Mammalian expression of zebra fish Oep with FLAG epitope tag inserted at N terminus following signal sequence | 67 |
| Nodal | ||
| pcDNA3-Nodal | Mammalian expression of mouse Nodal (coding region only; without epitope tag) | This work |
| pNodal/IRES-GFP | Generation of stable clones expressing mouse Nodal | This work |
| Type 1 receptors | ||
| pCMV5-ActRIB-HA | Mammalian expression of human ActRIB with HA epitope tag at C terminus | 2 |
| pCMV5-ActRI-HA | Expression of human ActRI with HA epitope tag at C terminus | 8 |
| pCMV5-BMPRIB-HA | Expression of human BMPRIB with HA epitope tag at C terminus | 32 |
| pCMV5-TβRI-HA | Expression of human TβRI with HA epitope tag at C terminus | 9 |
| Luciferase reporters | ||
| A3-lux | Luciferase reporter containing three tandem copies of Nodal and activin responsive element from the Xenopus Mix2 promoter | 31 |
| 3TP-lux | Luciferase reporter containing three tandem copies of TPA response element and part of PAI-1 promoter | 9 |
| (n2)-lux | Luciferase reporter containing seven tandem copies of Nodal-responsive element from the mouse Nodal promoter | 49 |