These phenotypes of the z3 mutant disappeared gradually and no distinct differences in phenotype were observed at 80 days after sowing, although the height of the z3 mutant was slightly shorter than that of the wild type, possibly due to its slower growth at the early stages (data not shown).
However we identified expression of EN and GsbN in NB 7-3 progeny at later stages (data not shown).
Interestingly, caspase-3 activation was only detectable during primitive hematopoiesis in these TERT morphants from 19 24 hpf, and not at later stages (data not shown).
This block was predominantly between the DP and single-positive (SP) stages (data not shown), which may explain why the tumors are arrested at the DP stage.
We did not find significant expression of ATF2 or ATF2-PT71 in other neurons of the brainstem at these developmental stages (data not shown).
We performed this assay in juvenile fishes (3 months old) since, in our hands, E2 does not induce a detectable luciferase response using these transgenic fishes at embryonic stages (data not shown).
We found that the addition of the WNT signaling inhibitor DKK1 only impacted cardiac differentiation when added at d0 d2 and none of the subsequent stages (data not shown).
Prior to HH24, expression of Scleraxis in head mesenchyme is general and diffuse, while Scleraxis expression clearly defines the syndetome domain within somites at the same stages (data not shown), [29].
However, although we see strong ubiquitous expression on the protein level for up to 7 days, our in situ analysis reveals strong ubiquitous expression only during gastrulation and mid-somitogenesis stages (data not shown).
Despite earlier reports [43], [55] that NT-3 mRNA can be detected by RT-PCR from before primitive streak formation, we were unable to detect it by in situ hybridization at these early stages (data not shown).
Reactivity was restricted to blood stages, since the antibodies did not react with sporozoites stages (data not shown), and this reactivity was also peptide-specific as shown by IFAT competition assays with the corresponding peptide (Figure 1A).
