Fig. 7. In vivo GSK3 injections perturb neural crest, neural plate and mesoderm development. (A-C) Injections in one of two-cell stage embryos of lacZ (A) or GSK3 (300 pg) plus lacZ mRNA (B,C). The GSK3 injected side (white arrow, pink lacZ staining) displays greatly reduced Slug expression (B, red arrow, blue staining), abnormal Krox20 expression (C, red arrow, blue staining) and reduction of the paraxial mesoderm marker 12-101 staining (B,C; black arrows, brown staining). Red and black arrows in A indicate normal staining for Slug (blue) and 12-101 (brown), respectively. (D-F) GSK3 (150 pg) was injected into one dorsoanimal blastomere at the 16-cell stage to target one neural fold and reduce the effect on adjacent tissues. (D) Control ß-galactosidase staining. (E) The injected area shows reduction in both Slug (red arrow) and 12-101 staining (black arrow). (F) Ectopic expression of the anterior neural plate marker Cpl-1 is induced ectopically (red arrow), showing that neural patterning is also affected.

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