XB-IMG-131089
Xenbase Image ID: 131089
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Fig. 8. Specificity of Cm-derrière.
(A) Rationale for dominant negative activity.
Solid bars and open bars indicate the prepro
region and the mature region of Derrière
protein, respectively. Gray boxes represent
mutated maturation cleavage signal. The
link between two open boxes represents the
disulfide bond of a dimer. (B) Experimental
scheme for C-E. Wild-type embryos were
injected at the animal pole in one cell at the
2-cell stage with test RNA. Animal caps
were removed from stage 8.5 (mid-blastula)
embryos and were cultured until sibling
embryos reached stage 20 (late neurula).
Animal caps and control whole embryos
were harvested for RT-PCR (C,D) and for
morphological analysis (E). (C) Ratio of
derrière: Cm-derrière at which marker
expressions are inhibited in animal caps.
Injection of globin alone served as a
negative control. ODC was used as a loading
control. Test RNA is indicated at the top. wt,
wild-type derrière; Cm, Cmderrière.
endodermin (Edd) is an endodermal
marker. Lanes as marked; lane 7, uninjected
whole embryo control. globin RNA was
injected at 1 ng; In derrière and Cm-derrière
injections 20 pg of derrière RNA was
injected with Cm-derrière RNA to make up
the indicated ratio of derrière: Cm-derrière;
and globin RNA making the total mass
injected equal to 1 ng. (D) Effects of Cmderrière
on other TGF-b family members:
molecular assay. Injection of Cm-derrière
alone served as a negative control. Test RNA
is indicated at the top and the presence or
absence of ten-fold mass excess of Cmderrière
is indicated by + or – sign. Lanes as
marked; lane 16, uninjected whole embryo
control. derrière, BVg1, Xnr1, Xnr2, Xnr3
and Xnr4 RNAs were injected at 50 pg and
activin RNA was injected at 5 pg.
(E) Effects of Cm-derrière on other TGF-b
family members: animal cap elongation
assay. Cm-derrière was used at a 10:1 ratio
to co-injected TGF-bs, shown above the
panels. derrière, BVg1, Xnr1, Xnr2, Xnr3
and Xnr4 RNAs were injected at 50 pg and
activin RNA was injected at 5 pg. Image published in: Sun BI et al. (1999) Copyright © 1999. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |