Fig. 3.

Condensation properties of Plk4 regulate centriolar copy number. a Amino acid sequence of mutation sites in Plk4. Red letters, mutation sites; Gray background, degron motif. b FRAP analysis of cytoplasmic GFP-Plk4 (Kinase + L1) condensates (b) and centriolar GFP-Plk4 (Full length) (c) in HeLa cells (n = 12 and 13 cells, respectively). For b and c, intensities were normalized with the average of three pre-bleach signals. Graph shows mean ± SD from two (b) or three (c) independent experiments. d Centriole over-duplication assay. Plk4 (Full length)-3xFLAG was exogenously expressed under the CMV mutant promoter in siPlk4-treated HeLa cells. Cells were transfected with siRNA (24 h) and then with plasmid (48 h). Cells were stained with anti-CP110 antibodies as a centriole marker. Scale bar, 10 μm; inset, 2 μm. Percentages of cells which have >4 CP110 foci were calculated. Graph represents mean percentages ± SD (siControl n = 146, siPlk4 + Emp n = 159, siPlk4 + WT n = 153, siPlk4 + 6A n = 153, siPlk4 + 7A n = 166, siPlk4 + 8A1 n = 155, siPlk4 + 8A2 n = 148, siPlk4 + 9A n = 151, siPlk4 + 10A n = 157, siPlk4 + KD n = 151 cells) from three independent experiments. e Summary of the correlation between the condensed Plk4 dynamics and frequencies of centriole overduplication. Source data are provided as a Source Data file