Fig 1: PLK1 inhibitors and an Aurora B inhibitor affect PLK1 localisation and the localisation of HEC1 and ZWINT-1 at kinetochores. Nocodazole-arrested cells were treated with either 1 μM GW843682X, 100 nM BI 6727, 500 nM Hesperadin or DMSO for 3 h in the presence of MG132 and nocodazole. Cells were stained with (A) anti-PLK1 (green), anti-ZWINT-1 (red) and CREST (blue) antibodies. Scale bar: 5 μm. (B) Representative image of Hesperadin-treated cells co-stained with anti-PLK1 (green) anti-CREST (red) antibodies and DAPI (blue). Scale bar: 5 μm. (C) Distribution of mitotic cells with large PLK1 foci after treatment with GW843682X, BI 6727, Hesperadin or DMSO as in A. At least 625 mitotic cells were counted for each treatment per repeat. (D) Cells were treated as in A and stained with anti-HEC1 (green), CREST (red) and DAPI (blue). Scale bar: 5 μm. (E) Cells were treated with nocodazole as above followed by shake-off and re-plated in the presence of nocodazole and 1 μM GW843682X, 100 nM BI 6727, 500 nM Hesperadin or DMSO for 3 h and in the presence of MG132. Proteins were analysed by western blot with the indicated antibodies.