Fig. 5. In ovo DNA electroporation into PSM. (A) A diagram showing positions of a plus electrode (platinum) under the embryo, and a minus electrode (sharpened tungsten) near the DNA solution which was laid over the anterior primitive streak at HH stage 7-8. (B) 24 hours after the electroporation with pCAGGS-GFP, GFP signal was observed in the segmented somites and PSM. (C) A sagittal section of a GFP-electroporated embryo (anterior to the left). (D) An embryo co-electroporated with pCAGGS-Lfng and pCAGGS-GFP showing no gross disturbance in the somites. (E) Western blotting showing that pCAGGS-Lfng and pCAGGS-NotchE, used for the electroporation, gave rise to proteins of expected size when transfected into COS cells. Products of Lfng/FLAG (42 kDa, lane 1) and NotchE/Myc (83 kDa of an unprocessed form, and 70 kDa and 63 kDa fragments of processed forms translocating to the nucleus (Kopan et al., 1996), lane 3) were, respectively, detected by anti-FLAG and anti-Myc antibodies. Lanes 2 and 4 are controls for lanes 1 and 3, respectively.

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