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Fig. 6. KD-Wee1 and YYY-FFF-Wee1 act as dominant-inhibitors to disrupt
gastrulation. The two dorsal blastomeres of four-cell embryos were injected
with 5-6 ng of RNA encoding ß-gal, KD-, YYY/FFF-, Shift- or Stop-Wee1 and
embryos were examined for blastopore formation at stage 11.5-12. Co-injection
of Wee1 and ß-gal (100 pg) and the subsequent staining for ß-gal
activity (red) shows the area of the embryo expressing the exogenous RNAs. (B)
RNAs used in A were analyzed by agarose gel electrophoresis and ethidium
bromide staining (top panel). Lysates prepared from embryos in A were examined
by immunoblot analysis using the FLAG antibody (middle panel). Percentage of
embryos with gastrulation defects (bottom panel). Number of embryos examined:
ß-gal (n=55), Stop- (n=78), Shift- (n=72), KD-
(n=82) and YYY/FFF-Wee1 (n=86).
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