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Figure 6


Fig. 6. MAPK signaling is required for transfating. Right panels show embryo whole mounts immunostained with mAb 6a9 and examined by confocal microscopy. The remaining panels show living embryos viewed with differential interference optics, except H and K, which are in situ hybridizations. (A-C) Control embryos at the late gastrula stage. 65-70 PMCs are arranged in a subequatorial ring pattern (arrow). (D-F) Embryos treated with 6 µM U0126 from the 2-cell stage. Arrows in E indicate locations adjacent to ectodermal thickenings where ventrolateral clusters of PMCs normally form. Arrow in F indicates one of two 6a9-positive cells that formed in this embryo, which was treated with U0126 until sibling controls reached the prism stage. (G-I) Control PMC-deficient embryos (not treated with U0126). Transfating is indicated by skeleton formation (G; 24 hours after surgery), Lvalx1 mRNA expression (H; 8 hours after surgery), and 6a9 immunostaining (I; 12 hours after surgery). (J-L) PMC-deficient embryos transferred to 6 µM U0126 immediately after surgery (J,K and L show embryos 24, 8 and 24 hours after surgery, respectively).





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