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Fig. 5. EZH2 and Snail2 regulate overlapping and distinct markers in Xenopus. (A) 10 pg Noggin and 25 pg Wnt8 RNAs, with or without 40 ng EZH2-MO or Snail2-MO, were injected into the animal regions of two-cell-stage embryos. Animal caps were dissected at blastula stage 9 and cultured until neurula stages 19-20. RNA was then extracted for RT-PCR analysis of marker expression. Co-expression of Noggin and Wnt8 induced a panel of neural crest markers in animal caps that were reduced by either EZH2 or Snail2 knockdown. Notably, however, depletion of EZH2 affected more markers (Pax3 and Zic1) than depletion of Snail2, indicating that EZH2 regulates a wider array of gene expression than Snail2. (B) 20 ng EZH2-MO or Snail2-MO was injected unilaterally into two-cell-stage embryos. The embryos were analyzed at neurula stages 16-17 by ISH for expression of Zic1 and Pax3. Knockdown of EZH2 reduced the posterior (arrow), but not the anterior (arrowhead), domain of Zic1 and the expression of Pax3; however, Snail2-MO was ineffective in reducing either Zic1 or Pax3. The numbers of embryos displaying the shown patterns over the total number of embryos analyzed are shown. |
