JCS008938 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Double-staining of multiple centrosome/cytoplasmic foci of GFP-centrin in HU-arrested cells with GFP-centrin (A-G) and Cep135 (A’-B’) or hCenexin (C’), pericentrin (D’), Nek2 (E’), ninein (F’) and PCM1 antibodies (G’). Merged images are shown in A”-G”. In cells undergoing multiple centrosome replications, centrosomal components localize differently. Bar, 10 μm.

• Supplemental Figure S2 -

  Fig. S2. Microtubule-nucleating activity of centrosomes/cytoplasmic foci of GFP-centrin in HU-arrested cells after brief recovery from nocodazole treatment (A,B). Cells were triple stained with Texas Red-conjugated monoclonal anti-α-tubulin and Cy5-conjugated polyclonal anti-γ-tubulin antibodies. Arrows indicate foci of GFP-centrin lacking γ-tubulin and microtubule asters. Bar, 10 μm.

• Movie 1 -

  Movie 1. Movement of the centrosome/cytoplasmic foci examined by time-lapse fluorescence microscopy. Images were acquired every 5 seconds.

• Movie 2 -

  Movie 2. Movement of the centrosome/cytoplasmic foci examined by time-lapse fluorescence microscopy. Frames at different time points of this movie are assembled in Fig. 7.

• Movie 3 -

  Movie 3. Movement of the centrosome/cytoplasmic foci examined by time-lapse fluorescence microscopy. The cell contains more cytoplasmic foci than those shown in Movies 1 and 2.

• Movie 4 -

  Movie 4. Inhibitory effect of nocodazole on the movement of centrosomes/cytoplasmic foci labeled by GFP-centrin. Cells were treated with 0.5 μg/ml nocodazole during the indicated period on the movie. After washing out nocodazole, the cells were further cultured in a fresh medium containing 2 mM HU.