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. Author manuscript; available in PMC: 2011 Feb 3.

Published in final edited form as: Cancer Cell. 2003 Apr;3(4):333–346. doi: 10.1016/s1535-6108(03)00083-7

A: Coimmunostaining of γ-tubulin (middle panels) and nucleophosmin B (right panels) in cells cultured under serum-starvation conditions.

B: Percentage of nucleophosmin B-positive unduplicated centrosomes in wild-type () or E2F3^−/− (■) cells under serum starvation conditions.

C: Percentage of cells with nucleophosmin B-positive centrosomes in the total population of wild-type () or E2F3^−/− (■) cells cultured under serum starvation.

D: Wild-type and E2F3^−/− cells expressing GFP-centrin-2. Shown from top to bottom: a wild-type cell with an unduplicated centrosome, prior to the separation of centrioles; an E2F3^−/− cell with a separated centrosome; an E2F3^−/− cell with multiple, separated centrioles. The left panels show the corresponding DAPI-stained nuclei. White arrows indicate individual centrioles while the red arrow indicates a separated centriole associated with a nascent centriole.

E: Top panel, percentage of wild-type () or E2F3^−/− cells (■) containing one (solid bars) or ≥2 separated centrosomes. Bottom panel, percentage of wild-type () or E2F3^−/− cells (■) containing unduplicated (solid bars), ≥2 separated and unduplicated (stripes), or ≥ 2 separated and duplicated centrosomes (dotted) in density-arrested conditions. Centrioles were visualized using GFP-centrin-2, and the calculated percentages are based on the results from counting 100 cells. Shown in parenthesis are the percentage of BrdU-positive cells in the population.

Inline graphic — A: Coimmunostaining of γ-tubulin (middle panels) and nucleophosmin B (right panels) in cells cultured under serum-starvation conditions.

B: Percentage of nucleophosmin B-positive unduplicated centrosomes in wild-type () or E2F3^−/− (■) cells under serum starvation conditions.

C: Percentage of cells with nucleophosmin B-positive centrosomes in the total population of wild-type () or E2F3^−/− (■) cells cultured under serum starvation.

D: Wild-type and E2F3^−/− cells expressing GFP-centrin-2. Shown from top to bottom: a wild-type cell with an unduplicated centrosome, prior to the separation of centrioles; an E2F3^−/− cell with a separated centrosome; an E2F3^−/− cell with multiple, separated centrioles. The left panels show the corresponding DAPI-stained nuclei. White arrows indicate individual centrioles while the red arrow indicates a separated centriole associated with a nascent centriole.

E: Top panel, percentage of wild-type () or E2F3^−/− cells (■) containing one (solid bars) or ≥2 separated centrosomes. Bottom panel, percentage of wild-type () or E2F3^−/− cells (■) containing unduplicated (solid bars), ≥2 separated and unduplicated (stripes), or ≥ 2 separated and duplicated centrosomes (dotted) in density-arrested conditions. Centrioles were visualized using GFP-centrin-2, and the calculated percentages are based on the results from counting 100 cells. Shown in parenthesis are the percentage of BrdU-positive cells in the population.