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. 2015 Jan 1;128(1):171–184. doi: 10.1242/jcs.163659

Fig. 5.

Fig. 5.

CENP-QS50A–eGFP rescues kinetochore recruitment of Plk1. (A) Immunofluorescence microscopy images (z-slice) of a CENP-QS50A–eGFP siRNA rescue experiment in HeLa K cells. Cells were treated with CENP-Q siRNA or control siRNA for 12 h and then transfected with an siRNA-resistant plasmid expressing CENP-QS50A–eGFP or a control eGFP expression plasmid for a further 48 h. To reduce the effect of the mitotic stage on alignment, cells were arrested in metaphase with 1 µM MG132 for 90 min before fixation. Cells were stained with CREST antisera (blue) and an antibody against Plk1 (red). Scale bar: 5 µm. (B) Quantification of Plk1 levels in the CENP-QS50A–eGFP siRNA rescue experiment. Staining intensities were determined at each kinetochore relative to that of CREST after background subtraction, n = 3, ≥300 kinetochores, 30 cells. The dashed line indicates CENP-Q levels in control-siRNA-treated cells. Error bars indicate ±s.d.