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Fig. 1. Cby RNA is supplied maternally and its level remains high throughout early Xenopus development (A) based on Xenbase data for X. laevis (red) and X. tropicalis (green); (B) RT-PCR analysis in X. laevis of Cby RNA at various developmental stages using ornithine decarboxylase (ODC) RNA as the normalizing control. In situ hybridization studies indicate that Cby is expressed at high levels in the neuroectoderm of stage 18 embryos (C); later in embryogenesis (D) Cby is expressed in a range of tissues including the myotome, pronephros, otic vesicle, central nervous system, migrating neural crest, the eye, and blood islands. Standard (E) and qPCR (F) analyses of ectodermal explants derived from control, Snail2/Slug, or Twist1 morpholino (MO) injected embryos revealed an increase in Cby RNA in response to inhibition of Snail2 expression. (G) Ectodermal explants were derived from embryos injected with GR-Snail2 RNA (200 pg/embryo) and either left untreated (–dex) or treated for 2 h with dexamethasone (+dex), dexamethasone and emetine (+dex, +eme), or emetine alone (+eme). Cby RNA levels were measured by qPCR with the Y-axis corresponding to the change in Cby RNA level with respect to control condition, either control MO injected (F) or in the absence of dexamethasone (G). Student t-test values of <0.05 are indicated by a “*”, while a p value<0.01 is indicated by “**” in this and all other figures. |
