PLK4-scaffolded foci that accelerate acentrosomal mitosis in TRIM37Δ cells are centrobin-independent. (A and B) Immunostaining and fluorescent signal of in situ–tagged CEP192-mNG in USP28Δ and TRIM37Δ RPE1 cells treated with DMSO or centrinone (A) and additionally treated with doxycycline to induce CNTROB or PLK4 knockout (B). Scale bars, 5 µm and 1 µm (insets). (C) Graph of the percentage of cells with condensates and CEP192-PLK4 foci in the indicated conditions. The experiment shown in A and B was performed twice with similar outcomes; data from one experiment are quantified in C. (D) Quantification of mitotic duration, measured as in Fig. 7 D, for the indicated conditions. Mean and 95% CI are plotted on top of the individual values; P values are from t tests. ****, P < 0.0001. The experiment was performed twice with similar outcomes; data from one experiment are quantified. (E) Images of acentrosomal spindles in centrinone-treated TRIM37Δ cells, with and without centrobin. The centrobin-scaffolded condensate (yellow arrow) and the PLK4-scaffolded foci (cyan arrowheads) are marked. Scale bar, 5 µm. The experiment was performed twice with similar outcomes. (F) Schematic summary of analysis of acentrosomal mitosis following centrinone treatment. The PLK4-scaffolded foci, and not the centrobin-scaffolded condensate, improve acentrosomal mitosis following centrinone treatment.