Fig. 13.

Centrosomal dynamics of GFP-CP224 and GFP-tubulin are largely unaffected by TACC depletion. Fluorescence recovery after photobleaching (FRAP) of GFP-CP224 and GFP-tubulin in TACC-RNAi cells was compared with FRAP in GFP-tubulin cells and GFP-CP224 cells, respectively, which express TACC at wild-type levels. In interphase cells whole centrosomes were bleached. For analysis of GFP-CP224 and GFP-tubulin dynamics during mitosis exclusively metaphase cells were chosen and a single spindle pole was bleached. GFP-recovery was followed by 4D confocal live cell imaging. Fluorescence intensities in the bleached region of interest were evaluated with ImageJ. Representative examples of time series of all cell types used for analyses were added as supplementary material. The respective GFP fusion protein, the cell type used and the cell cycle stage is depicted on top of the respective graph. The half time of recovery and the number of individual experiments combined are indicated on the lower right. The standard deviation is indicated as error bars