|Public on Jul 01, 2009
|Ectodermal explant after zic1 misexpression, 2nd biological replicate
|Neuralized ectodermal explants from zic1 and beta globin (control) injected embryos
|Neuralized ectodermal explants free from mesoderm
|Ectodermal explants were harvested from stage 9 embryos injected at the 2 cell stage with hormone-inducible zic1 or beta globin as control, each plus noggin to neuralize the explants. After incubation to the equivalent of stage 12 they were incubated with cycloheximide for 30 min to prevent further protein synthesis, followed by addition of dexamethasone to activate injected hormone-inducible zic1. Three hours later ectodermal explants were harvested and treated with proteinase K.
|RNA extracted with phenol/chloroform and purified by alcohol and sodium acetate precipitation
|Biotinylated cRNA were prepared according to the standard one-cycle Affymetrix protocol from 2 micrograms total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45C on GeneChip Xenopus laevis GeneChip Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|GeneChips were scanned using the Affymetrix GCOS Scanner 3000.
|gene expression of ectodermal explant expression of zic1 or beta globin control.
|Data was analyzed with Agilent Genespring GX v. 7.3.1 using Affymetrix GCOS generated flag data from scaled CHP files, and RMA-normalized CEL files.
|Jul 15, 2008
|Last update date
|Jul 18, 2008
|Montana State University
|Functional Genomics Facility
|109 Lewis Hall
|Expression data from ectodermal explants following activation of hormone-inducible zic1