Figure 8

Centrosome number in GFP-MNB/DYRK1A-overexpressing cells. HeLa cells were transfected with constructs encoding GFP, GFP-NLS, or MNB/DYRK1A as described in "Materials and Methods." The cells were then fixed, stained with anti-γ-tubulin antibody and TOTO-3, and observed by confocal laser scanning microscopy at the indicated time points. Each cell population (%) was determined by counting the centrosome number in each cell for a total of 200 cells. Paired centrosomes, which could be distinguished under microscopic observation, were scored as 2 centrosomes. The centrosome number was counted for non transfected cells (normal cells and those treated with LipofectAMINE but without a vector), for cells with fluorescence signal (GFP and GFP-NLS), and for cells with a diffuse distribution of fluorescence within their nuclei (GFP-MNB/DYRK1A). The data shown are means ± S.E. of 3 independent experiments.