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Display additional annotations [+]
| Gene |
Clone |
Species |
Stages |
Anatomy |
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fgf4
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laevis
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NF stage 18
to
NF stage 20
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involuted dorsal mesoderm
,
paraxial mesoderm
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fgf8
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laevis
|
NF stage 18
to
NF stage 20
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posterior neural tube
,
neural tube
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presumptive midbrain-hindbrain boundary
,
preplacodal ectoderm
,
anterior neural ridge
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cyp26a1
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laevis
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NF stage 18
to
NF stage 20
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mesoderm
,
axial mesoderm
,
paraxial mesoderm
,
posterior
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aldh1a2
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|
laevis
|
NF stage 18
to
NF stage 20
|
optic vesicle
,
lateral plate mesoderm
,
anterior dorsal lateral plate region
,
dorsal lateral plate mesoderm
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Fig. 7. The RA and Fgf pathways regulate each other. The levels of RA signalling (AI) were altered by addition of a synthetic RA antagonist (left column) or all-trans RA (right column) and compared to a DMSO control (centre column). Embryos were assayed for fgf4 (AC) and fgf8 (DI) expression. The posterior domain of fgf4 (a) is lost in RA treated embryos (C) when compared to the control (B), but unaffected in embryos treated with RAA. Expression of fgf8 is expanded both anteriorly (E and F) and posteriorly (H and I; compare distance between arrowheads (d) marking the anterior limits of domain, and (e) marking posterior limits of domain) under treatment with RA. Decreasing RA signalling also reduces the anterior domain of fgf8 underlying the heart region (compare ratio of staining intensity between (b) marking the pituitary anlagen to (c)). A similar effect is seen with sprouty2 expression (JO), as its domain is increased with RA in both the anterior heart region (L; arrowhead f) and it extends further anterior (g) in the dorsal neural tube (O) when compared to controls (KN). Conversely, embryos were treated with SU5402 and assayed for expression of aldh1a2 (P and Q) or cyp26 (R and S) to determine the effect of a loss of Fgf signalling on the RA signalling pathway. The expression domain of aldh1a2 was expanded posterior (Q) (arrowhead: h marking posterior limit of expression domain) as compared to control embryos (P). Cyp26, normally present in the posterior LPM tailbud domain (R; arrowhead i) is undetectable when Fgf signalling is inhibited (S). Ant: anterior view with dorsal at top of image. Dor: dorsal view with anterior at top. Llv: left lateral view with anterior toward left, dorsal at top of image. Pos: posterior view with dorsal at top. The total number of embryos examined for each panel is indicated in the lower left hand corner.
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Display additional annotations [+]
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Supplementary Fig. 2. Altering the bio-availability of endogenous RA ligand by inhibiting members of the RA signalling pathway leads to similar effects as exogenous RA treatments. Expression of cyp26, a biomarker of RA signalling, is slightly increased under treatment with ketoconazole (B and E), and decreased with DEAB (C and F) as compared to control embryos (A and D). The anterior fgf8 expression domain (arrowhead a) was unchanged with a moderate increase in RA signalling when treated with ketoconazole (H) was also reduced in comparison to expression in the pituitary anlagen (arrowhead b) under treatment with DEAB (I) when compared to controls (G). However consistent changes were seen with either ketoconazole or DEAB on the posterior domain of fgf8 expression (J–L). Also, no obvious and consistent changes were seen on the size and position of the sprouty2 domain (M–R). |
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Supplementary Fig. 4. Sprouty2 expression demonstrates the efficacy of SU5402. Treatment of embryos at stage 12.5 with 10 μM SU5402 with 0.1 mM ATP is sufficient to largely eliminate sprouty2 expression based on whole mount in situ hybridization when assayed at either stage 20 (A–D) or stage 32 (E F). |
