The Journal of Neuroscience, March 8, 2006, ():
RE-1 Silencer of Transcription/Neural Restrictive Silencer Factor Modulates Ectodermal Patterning during Xenopus Development
J. Neurosci. Olguín et al.
26: 2820
Supplemental data
Files in this Data Supplement:
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Figure S1. Alignment of the entire predicted amino acid sequence of rat, mouse human, Xenopus and zebrafish REST/NRSF.
Conserved identical residues are indicated by black boxes. The nine zinc fingers domains are boxed in red.
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Figure S2. XREST/NRSF mRNA expression pattern. XREST/NRSF is expressed in whole ectoderm at early gastrula stage, lateral view (a) and transverse section (b, arrowhead). During the early neurula stage, XREST/NRSF is restricted to the anterior and lateral neural ridge areas, anterior view (c, arrowhead ). At the neural tube stage, XREST/NRSF is expressed in the presomitic mesoderm, dorsal view (d, arrow) and transverse section (e, arrowhead). Later, it is detected surrounding the anterior nervous system and the developing eye (f, arrowhead) and slightly in the migratory neural crests (f, arrow) .
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Figure S3. β-galactosidase, control morpholine, and XREST1 injected embryos do not display altered expression of N-tubulin, Sox2 and Slug. Anterior views of stage 18 embryos injected with β-galactosidase mRNA, control morpholine and XREST1 mRNA (indicated on top). Probes used for ISH are indicated to the left of each row.
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Figure S4. Interference of XREST/NRSF function does not increases Notch signaling. Anterior-dorsal views of stage 16 embryos injected with 500 pg of dnXREST in one cell at the 2-cell stage. The activation of dnXREST at stage 9 results in the decrease of XHes-1 expression (arrowhead) (b), and apparently does not disturb Notch-1 expression pattern (d).
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Figure S5. Loss of XREST/NRSF function results in a persistent alteration of the patterning of the central nervous system. Anterior views of Xenopus larvae, showing the expression of Sox2 RNA. A control embryo is shown on the left; three embryos which were injected with 500 pg of dnXREST DNA in one cell at the 2-cell stage, and exposed to dexamethasone at stage 9 are shown to the right. Note the expansion of the Sox2-expressing domain in the side derived from the injected cell (shown to the right) and the loss of the normal contour of the eye field.
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Figure S6. The expansion of neural plate associated to the interference of XREST/NRSF function is rescued by the expression of zREST/NRSF.
Anterior-dorsal views of stage 18 embryos analyzed for Sox2 expression. Embryos were injected with MoXREST (a); MoXREST and 1ng of ZREST/NRSF (b); 200 pg of dnXREST/NRSF (c); 200 pg of dnXREST and 1ng of ZREST/NRSF (d). The size of neural plate is indicated with brackets and the anterior-lateral expansion is indicated with arrowheads (a, c). Coinjection of MoXREST and ZREST/NRSF RNA rescued the neural plate expansion of MoXREST injected embryos (a, b). Coinjection dnXREST RNA and the RNA encoding the full length ZREST/NRSF protein reversed the expansion of neural ectoderm caused by dnXREST alone (c, d).
