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Fig. 9. Char is localised at the inner nuclear membrane. (A,B)
Localisation of Char (red) and Lamin (green) in the NE of embryos viewed in
sagittal sections (A) and from the top (B). (C,D) WGA (green)
and Char (red) localisation at the NE of embryos viewed from the top
(C-C'') and in sagittal sections (D,D'). C shows a detail of
C' and D' a detail of D. Scale bars: 5 µm, except in C,D'
(300 nm). (E-H) Immunogold localisation of HA-Char (black
arrows) in an early embryo. The nucleoplasm (N) and cytoplasm (C) of three
different cells are indicated and have very different electron density. The
white arrows indicate contacting cell surfaces. HA-Char is concentrated at the
NE. High magnification views of representative localisation at the NE are
shown in F, where the white line defines the position of the NE. (G)
Representative localisation of Lamin. (H) Quantification of the localisation
of HA-Char and Lamin. We positioned the NE at the boundary between the
nucleoplasm and cytoplasm (white lines in F and G show examples) and
determined the localisation of gold particles (15 nm) at the boundary, or the
inner (in) or outer (out) side of the NE. Lamin and HA-Char are distributed
similarly, and are particularly biased towards the inner side of the NE.
(I,J) S2 cells stained with Char (green), Lamin (red), Hoechst
(blue) and Phalloidin (white) to mark F-actin at the cell cortex after
permeabilisation with Triton (I) or digitonin (J). Char staining is absent
from the NE when the plasma membrane but not the NE is permeabilised (with
digitonin).
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