Fig. 5. Over-expression of centrin-GFP disrupts the NBBCs. (A) Isolated cytoskeletons of control (a), Cen-GFP3 (b) and Cen-GFP8 (c) cells stained with anti-centrin (a'-c'), corresponding GFP images (phase contrast for the control) are shown in a-c. Over-expression of centrin-GFP disturbed the NBBCs (c), whereas moderate expression allowed incorporation into the proximal parts of the NBBCs and into the distal connecting fiber (arrow in b). Bar, 1 µm. (B) Dividing cells of Cen-GFP8 stained with anti-tubulin combined with either anti-centrin (a-c) or GT335 (d,e). (a-c) Small arrowheads, bb pairs; arrows, centrin-GFP clumps; large arrowheads, centrin/centrin-GFP located at the spindle poles. (d,e) Arrowheads, bb pairs detected by GT335 staining. Bar, 5 µm. (C) Phase contrast (a) and corresponding GFP image (a') of Cen-GFP8 cytoskeletons, which often disintegrated into basal apparatus (arrowhead) and cell nucleus (arrows). (D) Histogram showing the frequency of intact (solid bars) and defect (open bars) cytoskeletons. Data were averaged from three independent experiments. Cytoskeletons of the centrin-RNAi strain A7 and the centrin-GFP-overexpressing strain GFP8 were less stable.

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