Figure 3

Spindle pole aberrations in untransformed human mammary epithelial cells after paclitaxel treatment. (A) Double-immunofluorescence staining was performed to detect γ-tubulin (red) and α-tubulin (green). DNA was counterstained with Hoechst 33258 (blue). Panels show representative mitotic figures of CC cells with two (2SP), three (3SP), four (4SP) or five (5SP) spindle poles. Scale bar, 10 μm. (B) Representative time-lapse imaging of chromosomes in an untreated H2B-GFP expressing HME1 cell during metaphase–telophase transition. As indicated, cytokinesis was completed 24 min after metaphase (24:00). (C) Representative time-lapse imaging of chromosomes in a 1 nM paclitaxel-treated H2B-GFP expressing HME1 cell during metaphase–telophase transition. White arrows on 07:00 panel indicate the tripolar spindle orientation. Of note, 41 min (41:00) after metaphase, cell failed to enter telophase and irreversibly arrested in a metaphase-like arrest showing unaligned chromosome (white arrow). Time, mm : ss normalised to metaphase equals 00 : 00. Scale bar, 10 μm.