Fig. 4: AZD2811 treated cells have no colony forming or tumorigenic potential.

A Colony formation assays with the indicated cell lines in which cells were either untreated or treated for 3 days with AURKBi, then washed out, and plated to allow colony formation. B Similar experiment to (A) using immortalised human keratinocytes (EV) or EV cells expressing HPV E6/E7. C Nude mice were injected with either untreated or 5-day AZD2811 treated HCT116 RB^−/−p53^−/−. When the tumours were >100 mm³, the mice were culled, sacrificed, and examined upon autopsy. D Confocal microscopy of control C33A cells stained for DNA, γ-tubulin, α-tubulin to mark centrosomes and mitotic spindles and phalloidin for the actin cytoskeleton. E 6 day 200 nM AZD2811 treated mitotic cells. The bar in each is 10 μm. F C33A cell treated with 200 nM AZD2811 for the indicated times and stained as in (D, E) were visually inspected and centrosome number of the mitotic cells at each time point counted. G Parallel experiment to (F) but using either HCT116 wild type or RB^−/−p53^−/− cells. The data in (C) and (D) show mean and 95% confidence intervals.