Fig. 2.
Centromere clustering around the SPB is microtubule-dependent. (A) Combined IF/FISH images of G1-arrested cells probed with anti-p90 (SPB)-CY3 (red), anti-Nop1 (nucleolus)-CY5 (blue), and CEN3, CEN5, or CEN6 (green, as indicated). Full 3D rendering was performed with imaris (Bitplane, Zurich). Combined IF/FISH of diploid cells synchronized in G1 with combined probes to CEN8, CEN11, CEN13, and CEN14 (light blue), DNA stain (TOTO-3; blue), and anti-p90 (red). (Bar, 1 μm.) (B) Localization of GFP-tagged CEN4 relative to the nuclear periphery in G1-phase cells exposed to 1% DMSO with (filled bars) or without (open bars) 10 μg/ml nocodazole (noc). Time-lapse movies were acquired as in ref. 10. Relative 2D spot-periphery distances were averaged over six to nine movies (1,200–1,800 frames) and plotted in intervals of 0.2 μm (median value ± 0.1).