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Fig. 1. Antisense MOs against Bmp2, Bmp4 and Bmp7 inhibit endogenous Smad1
phosphorylation and cause dorsalization and posterior truncations of
Xenopus embryos. (A) Design of Bmp4, Bmp7 and Bmp2 antisense MOs that
target both pseudoalleles expressed in the subtetraploid species X.
laevis. (B) In vitro transcription/translation of Bmp4, Bmp7 and
Bmp2 is specifically inhibited by the respective MOs. (C) MOs for
Bmp2, Bmp4 and Bmp7 (at 12 ng each) were injected either alone or in
combinations at the four-cell stage radially in each blastomere. (D)
Endogenous carboxy-terminal Smad1 phosphorylation in stage 11 embryos is
decreased by co-injection of multiple Bmp MOs. (E-H) Bmp4 MO-injected embryos
(F) are dorsalized with enlarged heads (compare with control embryos, E). Red
arrowheads delineate the spinal cord marker Hoxb9 (>85%,
n=60). (G) Bmp4 MO-dorsalized phenotype is rescued by microinjection
of 100 pg of mouse Bmp4 mRNA. (H) Microinjection of mouse
Bmp4 mRNA alone (100 pg) results in ventralized embryos with small
heads, no eyes and reduced spinal cord structures (red arrowheads). (I-L)
Bmp4-depleted embryos (J) develop into swimming tadpoles with no ventral fins
and slightly larger heads than control embryos (I). Note the position of the
anus, which is displaced (posteriorized) to the tip of the tail (white
arrowhead; >72%, n=61). (K) Bmp7-depleted tadpoles develop with a
partial loss of ventral fin and a posteriorized anus (white arrowhead;
>79%, n=51). (L) Double knockdown of Bmp4 and Bmp7 results in
tadpoles lacking tail structures (>90%, n=39).
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