Fig. 6: cPlk1i-COOc allows spatial control of Plk1 activity in two and three-dimensional cell culture.

a Structure and uncaging reaction of cPlk1i-COOc. b Quantification of arrested cells in the experimental set up and conditions shown in Fig. 5d, e. Cells were treated with DMSO or cPlk1i-COOc (1 µM). DMSO control is the same as in Fig. 5e and Supplementary Fig. 10 as both inhibitors were tested in the same experiments. Graph shows mean ± SD. Number of cells n = 4037, N = 2. Each condition was compared to the dark DMSO control of the respective condition by performing ordinary one-way ANOVA. **: p = 0.0033, ns: p ≥ 0.999 (exact p values are provided in the Source Data file). c Scheme of the experimental setup in spheroids. d Confocal images of consecutive time frames of stably expressing H2B-mCherry (black) spheroids treated with DMSO or cPlk1i-COOc (5 µM). The pink dotted line box on the left highlights the illuminated quadrant of the spheroid, as shown in the scheme in panel c, the cyan dotted line box on the right marks the dark quadrant. The smaller solid-line boxes within the illuminated and dark quadrants represent examples of individual cells. Magnified views of the solid-line boxes are shown on the right of the spheroid. Time points indicated in hh:min. Scale bar 50 µm, scale bar in the 5x zoom 5 µm. e Cumulative frequency plot of mitotic duration. The dotted or full lines show the mean, the shaded areas are the SD, n = 983, N = 3. The Mann-Whitney test was used to compare medians (Supplementary Table 1). Source data are provided as a Source Data file.