Xenbase Image ID: 82119

Fig. 8. Induction of XPax-2 splice variants in animal cap explants. Embryos were cultured until stage 8, at which the upper part of the animal hemisphere (animal cap) was removed. The animal caps were treated for 12 h at 22 with either activin (10 ng/ml), retinoic acid (RA, 10 mM), basic FGF (bFGF; 100 ng/ml) or a combination of activin and RA. Control explants were cultured in the absence of any factors. Expression of XPax-2 splice variants was assayed by RT-PCR (panel A-C) and by whole mount in situ hybridization (panel D) once reference embryos reached stage 28. In panels A and B, schematic representations of the XPax-2 protein are shown to illustrate the RT-PCR strategy. Autoradiographs is shown with the size markers on the left. (A) Identification of alternative splice variants using primer pair XP-59/XP-63. (B) Identification of alternative splice variants using primer pair XP-9/XP-24. (C) Control for equal RNA amounts. RT-PCR experiments were carried out in parallel with elongation factor-1a (EF-1a) specific primer pair EF1a-1/ EF1a-2. Amplification products were separated by agarose gel electrophoresis and visualized by staining with ethidium bromide. M, lane with size markers. (D) Induction of XPax-2 expressing tissues in animal cap explants. Animal caps were incubated either under control conditions or in presence of added factors (activin alone; activin and RA) as described above. Note the absence of XPax-2 transcripts in control animal caps. Image published in: Heller N and Brändli AW (1997) Copyright © 1997. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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