DEAB rescue of cdx4−/− embryos requires exposure during gastrulation
| Stage exposed . | Time exposed, hpf . | Mutant embryos with increased gata1/total mutants . | Mutant embryos with increased gata1, % . | Degree of rescue . |
|---|---|---|---|---|
| None | N/A | 0/23 | 0 | None |
| 50%-90* | 5.5-9 | 12/19 | 63 | 2+ |
| 50%-1 ss | 5.5-10.3 | 23/24 | 96 | 2+ |
| 90%-1 ss* | 9-10.3 | 18/21 | 86 | 1+ |
| 1 ss-5 ss* | 10.3-11.7 | 2/25 | 8 | Scant |
| 50%-5 ss | 5.5-11.7 | 21/23 | 91 | 3+ |
| 60%-10 ss* | 6-14 | 8/8 | 100 | 3+ |
| 90%-5 ss | 9-11.7 | 9/9 | 100 | 2+ |
| 90%-10 ss | 9-14 | 10/10 | 100 | 1+ |
| 5 ss-10 ss* | 11.7-14 | 0/6 | 0 | None |
| Stage exposed . | Time exposed, hpf . | Mutant embryos with increased gata1/total mutants . | Mutant embryos with increased gata1, % . | Degree of rescue . |
|---|---|---|---|---|
| None | N/A | 0/23 | 0 | None |
| 50%-90* | 5.5-9 | 12/19 | 63 | 2+ |
| 50%-1 ss | 5.5-10.3 | 23/24 | 96 | 2+ |
| 90%-1 ss* | 9-10.3 | 18/21 | 86 | 1+ |
| 1 ss-5 ss* | 10.3-11.7 | 2/25 | 8 | Scant |
| 50%-5 ss | 5.5-11.7 | 21/23 | 91 | 3+ |
| 60%-10 ss* | 6-14 | 8/8 | 100 | 3+ |
| 90%-5 ss | 9-11.7 | 9/9 | 100 | 2+ |
| 90%-10 ss | 9-14 | 10/10 | 100 | 1+ |
| 5 ss-10 ss* | 11.7-14 | 0/6 | 0 | None |
Embryos from incrosses of cdx4+/− heterozygotes were treated with DMSO or DEAB at various stages of early development. (5.5 hpf = 50% epiboly, 6 hpf = 60% epiboly, 9 hpf = 90% epiboly, 10.3 hpf = 1 ss, 11.7 hpf = 5 ss, and 14 hpf = 15 ss) The total number of mutant embryos in 2- to 3-treated groups were pooled for each incubation period. Embryos were incubated in DEAB (2 × 10−5M) and compared with control embryos incubated in DMSO to evaluate the degree of gata1 rescue. Each data point represented pooling of several (2-4) experiments. The degree of rescue was scored based on the number of gata1+ cells present in comparison to control embryos for that experiment. A full rescue was scored as 3+ and reflected a maximal increase of gata1+ cells as depicted in Figure 4D. When only a few more cells were present in comparison to the vehicle-treated control embryos, these embryos were scored as 1+ rescue. The level of rescue was deemed intermediate or 2+ if the number of gata1+ cells was less than maximal, or if there were mixed levels of rescue among the total number of mutant embryos scored.
N/A indicates not applicable.
Treatment groups depicted in Figure 5.