Smad1 rescues the Smad5 mutant or morphant, but Smad5 does not rescue the Smad1 morphant
| Mutant or morphant . | (Co-) Injected with . | n . | Phenotype at 24 and 48 hpf, % . |
|---|---|---|---|
| Smad1 MO | Smad1* RNA | 109 | 69 normal, 31 overexpression phenotype† |
| Smad5 MO | Smad5* RNA | 120 | 93 normal, 6.6 morphant phenotype |
| Smad1 MO | Smad5 RNA | 300 | 100 morphant phenotype |
| Smad5 MO | Smad1 RNA | 160 | 79 normal, 21 morphant phenotype |
| pgy/+ cross | Smad5 RNA | 200 | 100 normal |
| pgy/+ cross | Smad1 RNA | 205 | 78 normal, 1 pgy/pgy phenotype, 21 overexpression phenotype† |
| Mutant or morphant . | (Co-) Injected with . | n . | Phenotype at 24 and 48 hpf, % . |
|---|---|---|---|
| Smad1 MO | Smad1* RNA | 109 | 69 normal, 31 overexpression phenotype† |
| Smad5 MO | Smad5* RNA | 120 | 93 normal, 6.6 morphant phenotype |
| Smad1 MO | Smad5 RNA | 300 | 100 morphant phenotype |
| Smad5 MO | Smad1 RNA | 160 | 79 normal, 21 morphant phenotype |
| pgy/+ cross | Smad5 RNA | 200 | 100 normal |
| pgy/+ cross | Smad1 RNA | 205 | 78 normal, 1 pgy/pgy phenotype, 21 overexpression phenotype† |
Wild-type embryos were injected with the indicated combinations of MO, and capped RNAs or mutants were injected with capped mRNA at the 1-cell stage. The established doses of MOs were used (2 ng Smad1 MO, 1 ng Smad5 MO). The following doses of RNA were used for injection: Smad1*, 450 pg; Smad5*, 300 pg; Smad5, 75 to 300 pg; and Smad1, 75 to 150 pg. Each dose was titrated to avoid an overexpression phenotype and provide optimal rescue. The phenotypes were then scored at 24 and 48 hpf for gross morphology by microscopic examination. The following phenotypes were seen: normal (appearing wild-type), Smad1 overexpression (as described27 ), smad5 morphant (phenocopies pgy/pgy; Figure 2A; text), smad1 morphant phenotype (Figure 2A; text), and pgy/pgy mutant (Figure 2A). The number of embryos scored (n) and percentage displaying each phenotype was tabulated as shown.
Smad1 overexpression phenotype as described by Dick et al.27