Fig. 7. Cells overexpressing GFP-C-Nap1 establish functional mitotic spindles. (A) GFP-C-Nap1 expression was induced for 24 hours and cells were stained for MTs in red (anti--tubulin mAb), DNA in blue (Hoechst dye) and C-Nap1 in green (GFP fluorescence). Representative cells are shown in interphase (a), metaphase (b) and anaphase (c). Bars, 10 µm. (B) Further immunofluorescent analysis of GFP-C-Nap1-expressing cells. Staining in red shows poly-glutamylated tubulin that predominantly localizes to centrioles (GT335; a,b) and spindle poles (anti--tubulin; c); staining in blue shows DNA and in green shows C-Nap1. Shown are representative cells in interphase (a) and mitosis (b,c). Bar, 10 µm. (C) Prophase cells were stained in red for -tubulin (a) and -tubulin (b), in blue for DNA and in green for C-Nap1. Enlarged (2x) insets of the C-Nap1 patches surrounding the centrosome are shown in black and white (b); z₀ and z₁ designate two different focal planes. Bar, 10 µm.

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