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Figure 3. Twist and Cxcr4, but not Snail1 or Snail2, are Hif-1α targets in the neural crest. (A–L) Xenopus embryos. In situ hybridization for Snail1, Snail2, Twist, and Cxcr4 after ATGMOhif-1α injection. Asterisks show the injected side. (A–F) Snail1 and Snail2 expression. Note that neither Snail1 nor Snail2 are affected by inhibition of Hif-1α, but a clear inhibition of neural crest migration is observed. (G–I) Twist expression is lost after ATGMOhif-1α injection (arrows). (J–L) Cxcr4 expression. (J) Note that strong expression in the neural plate is not affected, whereas the neural crest expression is visible only in the right (noninjected) side as three masses next to the neural plate. Arrows indicate absence of gene expression in the neural crest at the injected side. (M and N) Quantification of results shown in A–L. Bars represent the standard deviation from three independent experiments, a minimal of 40 embryos was analyzed for each case. (O and P) Quantitative PCR (O) and RT-PCR (P) of neural crest dissected from embryos injected with a control MO or with ATGMOhif-1α genes analyzed as indicated. EF-1α was used as a loading control. Bars represent the standard deviation from three independent experiments. A minimal of 30 embryos was analyzed for each case. ***, P < 0.005. |
