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-CharCSIM) 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.