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Fig. 8. Char is farnesylated. (A,B) HA-Char localises to the
nuclear envelope in a cellularising embryo (A), whereas a Char mutant protein
deleted of the farnesylation motif CSIM (HA-Char
CSIM) is mostly present
in the cytoplasm and the nucleoplasm, although traces are detected at the NE
(B). (C) Western blot showing the different migration on SDS-PAGE of
HA-Char and HA-CharCSIM from the lysate of Drosophila S2 cells
in the absence or presence of 10 to 40 µM of the farnesyl-transferase
inhibitor FTI-277. The arrow indicates the position of the fast migrating,
non-farnesylated fraction of Char (lower band). (D) Injection of
FTI-277 60 minutes prior to cellularisation causes a `char-like'
phenotype: the nuclei round up and fall from the cortex. The inset shows a
detailed view of the boxed area. The nuclei (in white) are not properly
anchored apically (arrows). (E) Confocal section from the top showing
the nuclear morphology and position with Hoechst (green) and Lamin (red).
z-stack projections are shown at the top and to the right showing the
abnormal positions of the nuclei viewed from the side. (F) In
FTI-injected embryos during cellularisation (right), Char is no longer present
at the NE compared with control water-injected embryos (left), whereas Lamin
localisation is not affected yet.
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