|
Display additional annotations [+]
| Gene |
Clone |
Species |
Stages |
Anatomy |
|
sebox
|
|
laevis
|
NF stage 11
|
mesoderm
,
marginal zone
,
involuting marginal zone
,
blastopore lip
,
involuted dorsal mesoderm
,
[+]
|
|
bix1.3
|
|
laevis
|
NF stage 11
|
mesoderm
,
marginal zone
,
involuting marginal zone
,
involuted dorsal mesoderm
,
involuted ventral mesoderm
,
[+]
|
|
| |
Fig. 2. Activation of sebox expression by Nodal/Activin signaling. A: WISH in mid-gastrula stage (st11) embryos injected with 80 pg Xnr1 mRNA
into one cell at the four-cell stage and analyzed for sebox and brachyury (bra) expression. All embryos were viewed from the lateral side (lat.) with
the animal pole upward. B: qRT-PCR analysis of the expression of sebox, bra, goosecoid (gsc) and chordin (chrd) in animal caps (ACs) in response
to Activin treatment. Animal caps were dissected at st8.5 and cultured with the indicated dose of Activin for 4 hr before being subjected to RTPCR
analysis. The gene expression levels in the control whole embryos (WE) were set to 100% for normalization and odc served as a loading
control. C: WISH of st11 embryos injected with 2 ng cerberus-short (Cer-S) mRNA at the two-cell stage and analyzed for sebox and bix4 expression.
Embryos were viewed from the vegetal (veg.) pole with the dorsal side upward. D: qRT-PCR analysis of the relative expression of the indicated
genes in mid-gastrula (st11) embryos with treatments, as shown in C. The expression level of odc was used for normalization. E: RT-PCR
analysis of the expression of the indicated genes in animal cap explants at the equivalent stage 11. Animal caps were isolated from st8.5 embryos
that were injected with 0.5 ng VegT mRNA with or without 2 ng DN-ActRII mRNA at the two-cell stage. The marker odc served as a loading control.
(F) RT-PCR analysis of the expression of the indicated genes in st11 animal cap explants that were isolated from st8.5 embryos injected with
a range of doses (0–300 pg) of bra mRNA. The marker odc served as a loading control. |
|
Display additional annotations [+]
|
| |
Fig. 6. Sebox is required for the expression of Cerberus. A: Embryos at stage 12 were collected for WISH for pcdh8.2. Scale bar¼1 mm. B: Histogram
presentation shows the mean width of the gap between the bilateral pcdh8.2 expression domains. ***P<0.001 by Student’s t-test. C,D:
Transversely sectioned view of embryos showing pcdh8.2 (C) or bra (D) mRNA expression. A total of 80 ng Sebox MO was injected into the marginal
zone of both cells at the two-cell stage. Red boxes in C demarcate the notochord (nc). E: Embryos at stage 11 showing mRNA expression
of cerberus (cer) or gsc. Sebox MO (80 ng), alone or combined with rescuing sebox mRNA (100 pg), was injected into both cells at the two-cell
stage. All embryos were viewed form the dorsal side with animal pole upward. F: RT-PCR analysis of the expression of cer, gsc, sizzled (szl), and
gata5 in sebox-overexpressing ventral marginal zone explants (VMZ). VMZ or DMZ from control embryos or embryos injected with increasing
doses of sebox mRNA (0, 50, 100, and 200 pg) were dissected at st10.25 and collected at st11 for RT-PCR analysis. The marker odc served as a
loading control. G,H: Embryos at stage 26 (G) or stage 41 (H). A total of 80 ng control or Sebox MO was injected into the marginal zone of both
cells at the two-cell stage. The red solid lines in G were drawn to measure the size of the head. The encircling lenses and the coiling gut in the
embryos shown in H are indicated by red arrows and red asterisks, respectively. I: Histogram presentation of the relative head size of the
embryos, measured by drawing a line from the cement gland to the midbrain area in the control or Sebox-depleted embryos, as indicated in G.
**P<0.01 by Student’s t-test. |
