Fig. 10. The C-terminal domain of NUANCE targets the NE. (A-C) Confocal optical
section through the apical part of the COS7 nucleus showing that the
overexpressed GFP-Cterm1 protein was arranged in patches displacing nuclear
pores (arrows), detected by anti-Nup358 antibody (B). (D-F) The GFP-Cterm2
remained associated with the NE in prophase (D, arrow) when the nuclear pores
begin to disassemble as indicated by diffuse anti-Nup358 staining (E). DNA was
visualized with DAPI (F,I,L). (J-L) Two examples of the cellular localization
of the GFP-NUA
460-6643 overexpressed in COS7 cells stained
with TRITC-phalloidin (H,K). The GFP-NUA
460-6643 possesses
the N-terminal ABD and the C-terminal Klarsicht-like domain but lacks most of
the central rod domain. The GFP-NUA
460-6643 is arranged in
fibers (G-I, small arrowheads) or perinuclear aggregates (J-L, arrows)
partially colocalizing with actin. Dynamic cortical cytoskeleton (G-I, arrow)
and some of the stress fibers (G-I, large arrowhead) do not recruit
GFP-NUA
460-6643. (I,L) show overlays of the GFP, TRITC and
DAPI channels. Bars (A-C) 5 µM; (D-F) 10 µM; (J-L) 20 µM.