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. 2021 May 28;4:640. doi: 10.1038/s42003-021-02158-2

Fig. 4. AURKA remains active at centrosomes upon PROTAC treatment.

Fig. 4

a–c Immunofluorescence analyses of U2OS cells synchronized through mitosis and treated for 3 h with MLN8237, PROTAC-D, or DMSO vehicle control. AURKA staining is retained at the centrosomes after PROTAC-D treatment (a) whilst a marker for active AURKA at centrosomes, p(Ser83)LATS2, persists after PROTAC-D treatment but not MLN8237 treatment (b, c). c Measurement of centrosomal pLATS2 staining from immunofluorescence images after cytoplasmic background subtraction (Kruskal–Wallis multiple ANOVA, and Dunn’s post-hoc multiple comparison test to DMSO for significance). d, e Mitotic phenotypes scored from TPX2/DAPI staining of fixed cells (categories illustrated in Supplementary Fig. S3). d Dose-dependence of phenotypes using pooled data from three coverslips for each condition from a single experiment. e Mitotic categories scored at a single dose (100 nM), using data from two coverslips each from three independent repeats of the experiment, mean values ± SDs for each experiment are plotted. f Line-scans through fixed mitotic cells stained with pLATS2 (as shown in b), to show both poles of the spindle (visible as maxima of staining intensity), reveal reduced pole–pole distance after PROTAC-D treatment. Traces show mean values from n ≥ 22 individual mitotic spindles, with fainter traces above and below representing SDs.