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Figure 3 | SpringerPlus

Figure 3

From: p31comet inactivates the chemically induced Mad2-dependent spindle assembly checkpoint and leads to resistance to anti-mitotic drugs

Figure 3

Localization of Mad2 and microtubule arrays of in p31 comet -overexpressing cells in the presence of anti-mitotic drugs. (a) Mad2 localization in p31comet overexpressing cells. HeLa cells stably expressing EGFP-Mad2 were infected with EGFP or EGFP-p31comet adenovirus for 24 h, washed with fresh medium twice, and treated with anti-mitotic drugs at the indicated concentration for 24 h described in Figure 2. Cells were fixed with 3% paraformaldehyde and counterstained with Hoechest33324 for DNA (shown in blue). EGFP-Mad2 was shown in green. Cells were treated with the following drugs: (upper panels) nocodazole, (middle panels) taxol, and (lower panels) monastrol. EGFP (left panels) or EGFP-p31comet-expressing (right panels) cells were shown. (b) Array of microtubules in p31comet-overexpressing cells. HeLa cells were infected with EGFP or EGFP-p31comet adenovirus for 24 h, washed with fresh medium twice, and treated with anti-mitotic drugs at the indicated concentration for 24 h. Cells were fixed with cold methanol, stained with β-tubulin antibody for microtubules (shown in green), and counterstained with Hoechest33324 for DNA (shown in blue). Cells were treated with the following drugs: (upper panels) nocodazole, (middle panels) taxol, and (lower panels) monastrol. EGFP (left panels) or EGFP-p31comet-expressing (right panels) cells were shown. Note: EGFP and EGFP-p31comet signals were not detectable in this condition. (c) Array of microtubules and Mad2 localization in p31comet overexpressing cells. HeLa cells were infected with EGFP-p31comet adenovirus for 24 h and, treated with Taxol for 24 h. Cells were fixed with cold methanol, stained with Mad2 (upper panel) or α-tubulin (lower panel) antibodies (shown in green), and counterstained with Hoechest33324 for DNA (shown in blue).

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