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Fig. 3. Zebrafish dpr1 and dpr2 morphant phenotypes. (A) RT-PCR
analysis of efficacy of dpr1 MO. dpr1 splice blocking MO
induces the predicted shift to an 
1700 band, indicating it abrogates
proper splicing of the transcript. Spliced band is at 500 bp. U,
uninjected; I, injected. (B) 5'-Dpr2-luciferase construct is blocked in
an in vitro transcription/translation reaction. ß-Galactosidase was used
as an internal control. (C) dpr1 morphants (b, injected with 12
ng MO) are discernibly smaller but have no major change of cell fates compared
with control (a). dpr2 morphants (c, injected with 8 ng MO) have
convergent extension defects, which are more apparent in whole embryos (D,
parts c,d). However, the notochord is noticeably wider – compare
distances between the small posterior arrowheads in c with a and b.
dpr1+dpr2 morphants (d) do not have novel phenotypes,
indicating they are not redundant. White arrow indicates anterior limit of
head. Black arrows indicate distance between opl and en2.
Small arrowhead indicates somite. Posterior arrowheads indicate width of
notochord. Embryos were flatmounted, with anterior towards the left. (D)
Whole-mount in situ hybridizations of dpr2 morphants indicate they
are shorter (compare arrowheads in a and c) and the somites and notochord are
wider (compare myod staining in b and d), but major specification
events are not affected. For C and D, the experiments were repeated four times
with comparable results, see text for penetrance of phenotypes. In situ probes
used from anterior to posterior were: cathepsinL
(catL-hatching gland), opl (telencephalon), en2
(midbrain/hindbrain boundary), krox20 (rhombomeres 3 and 5) and
myod (adaxial cells and somites).
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