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. Author manuscript; available in PMC: 2020 Jan 17.
Published in final edited form as: ACS Omega. 2019 Dec 24;5(1):822–831. doi: 10.1021/acsomega.9b03626

Figure 4.

Figure 4

Treatment with compound (-)-9 causes mis-localisation of PLK1 from kinetochores in mitotic cells. (A) Cells expressing GFP-PLK1 were treated with compound (-)-9 or DMSO for 9.5h after double thymidine release. The cells were fixed, stained for DNA (Hoechst) and kinetochores (CREST anti-sera) and analysed by immunofluorescence microscopy. Pro-metaphase mitotic cells were identified based on DNA morphology and 1μm confocal Z-stacks were taken for each cell. Quantification of GFP-PLK1 intensity on CREST-stained kinetochores was carried out using ImageJ. The data is represented as Whisker maximum to minimum plot with horizontal bar indicating mean GFP-PLK1 intensity on kinetochores. Statistical analysis was done using non-parametric, Mann-Whitney two-tailed test with 95% confidence interval, **p=0.0037. 8 cells were analysed for DMSO and 7 for Compound (-)-9. (B) Representative maximal-intensity projection images of cells showing CREST-stained kinetochores (red), GFP-PLK1 (green) and DNA (blue) used for quantification of GFP-PLK1 intensity in Fig. 7A. Scale bar is 5μm.