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Fig. 7. Nucleolin depletion causes mitotic spindle defects. (A) Immunoblot showing effective depletion of nucleolin and phospho-nucleolin after nucleolin RNAi. ${alpha}$ -tubulin is shown as a control. (B) Mitotic cells stained for phosphorylated nucleolin (green). (C) Mitotic cells stained for phosphorylated nucleolin (green) and ${gamma}$ -tubulin (red). (D) Mitotic cells stained for B23 (green). Arrowheads show fluorescence signals for B23 at the spindle poles. (E) Mitotic spindle defects in nucleolin-depleted cells. The majority of cells with chromosome congression defects (i, misalignment; ii, non-alignment) appear to have normal spindles, whereas a small percentage show disorganized (iii and iv) or multipolar (v) spindles. Staining for nucleolin (green) and ${alpha}$ -tubulin (red) is shown. (F) Histograms showing quantification of the spindle defects after nucleolin depletion. Values represent the mean of four independent experiments ± s.d. (n=400). (G) Nucleolin depletion induces multiple centrosomes. Staining for ${gamma}$ -tubulin (green) and ${alpha}$ -tubulin (red) is shown. Bars, 5 µm.

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