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Fig. 6. X-TSK interacts with the Notch signaling pathway.
Whole-mount in situ hybridization analysis of mRNA-injected embryos. Injected
samples are indicated in the upper right-hand corners; probes are indicated in
the lower right-hand corner. (A,C) Neural crest markers Xslug
(A) or Sox9 (C) disappeared or diminished (arrowheads) with
X-TSK (1 ng) overexpression. (B) The ß-gal mRNA (0.5 ng)
injection had no effect. (D) X-TSK-CD2 mRNA (1 ng) showed a
similar effect to that in C. (E) Sox2 expression expanded
laterally (right bar) in the X-TSK (1 ng)-injected side. (F)
Epidermal keratin levels diminished. The anterior border disappeared on the
injected side (open arrowhead). (G) An embryo injected with
Chordin (100 pg). The Sox2-expressing neural plate is
expanded (right bar). (H) An embryo injected with Chordin (100
pg). Expression of the neural crest marker Sox9 diminished
(arrowhead). (I,J) X-TSK-si (0.5 pmol) and Chordin mRNA (100
pg). Faint expression of Sox9 (I) or Xslug (J) was observed
in the far lateral side (arrowheads). (K) Hairy2A expression
is downregulated on X-TSK-injected side, though it appears to spread
over a broader domain (arrowhead and open arrowhead). (L)
X-TSK mRNA (1 ng) injection resulted in decreased expression of
Msx-1 (arrowhead) though the width of its expression area (right
bracket) becomes wider than in the control side. (M) An embryo injected
with X-TSK (1 ng). Xbmp4 expression increased outside the
injected side (arrowhead). (N) An embryo injected with Chordin
(100 pg). Xbmp4 expression disappeared around the ß-gal-positive
cells (arrowhead). (O) BMP4 (0.5 ng) mRNA and (P)
Notch ICD (0.5 ng) injections caused reduction of Xslug
expression (arrowheads). (Q) ß-Gal mRNA injection did not affect
Xbmp4 expression. (R) X-delta-1Stu
injected areas are marked by ß-gal staining (light blue), the arrowhead
indicates ectopic Xbmp4 expression (dark purple). (S,T) An
embryo injected with X-TSK. (S) Two out of the three strips of
N-tubulin expression disappeared (arrowheads). (T) X-delta-1
expression disappeared on the outer border of the neural crest (arrowhead).
(U) ß-gal did not affect X-ESR-1 expression. (V)
X-TSK mRNA (1 ng) injection resulted in expanded peripheral
expression of X-ESR-1 on the injected side (arrowheads). (W,X)
Co-injection of 1 ng X-TSK and 0.5 ng Su(H)DBM
mRNAs. (W) Diffused signals of Sox9 were observed around the
arrowhead. (X) Xbmp4 expression was extended towards the
ß-gal-positive side (arrowhead). (Y) Sox9 expression was
reduced in Su(H)/Ank 500 pg injected side (arrowhead). (Z)
Sox9 expression was completely missing in X-TSK 1 ng+
Su(H)/Ank 500 pg injected side (open arrowhead). (AA)
Xbmp4 expression was very weakly inhibited in Su(H)/Ank 500
pg injected side (open arrowhead). (BB) The patchy expression of
Xbmp4 was observed in X-TSK 1 ng+ Su(H)/Ank 500 pg
injected side (arrows). The expansion of Xbmp4-expressing regions was
not observed (open arrowhead).
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