Fig. 6. Localization of NUANCE compared with that of nuclear pores. Cells were double labelled with anti-NUANCE (A,D,G,J) and anti-nucleoporin358 (Nup385) (B,E,K) antibodies. A-C, confocal sections of the apical part of a COS7 cell nucleus. The optical surfaces of nuclei are shown with enlargements of the regions indicated. NUANCE- (arrowheads, A-C) and Nup358-positive spots (arrows, A-C) are only partially colocalized. (D-L) The equatorial optical sections from the nuclei of COS7 cells, treated with 0.5% Triton X-100 (D-I) and 4 µM digitonin (J-L). Visualization of NUANCE at the NE in digitonin-treated cells (J-L, arrowheads) suggests its localization on the cytoplasmic face of the ONM. Note the polar distribution of cytoplasmic NUANCE (D, J, asterisk) and Nup385. In Triton X-100-treated cells, the intranuclear labeling was enriched in the nucleoli (D-I, arrows), which was verified by anti-NO38 staining (H). Bar, 5 µm.

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