Figure 4: Malignant T-cells undergo anaphase catastrophe following NAE inhibition (IMAGE)
Caption
(A) Cells were incubated with the indicated concentration of pevonedistat or vehicle control for 24 hours. Cells were immunostained with γ-tubulin (pink) and counterstained with DAPI (blue). Total anaphases were counted (50) and the proportion of multipolar anaphases is shown. Data are the mean ± SEM of at least 3 independent experiments. Representative examples of normal mitosis and multipolar anaphase are shown. (B) Jurkat and SR cells were transfected with a lentiviral CRISPR toolbox to achieve knockout of p27, or vector control. Cells were treated with pevonedistat as indicated, proteins were lysed and subjected to immunoblotting. (C) Cells were treated with pevonedistat for 24 hours and subjected to cell cycle profiling with propidium iodide staining. A summary of three independent experiments is shown. (D) p27-deficient cells were incubated with 0.25 μM pevonedistat or vehicle control for 24 hours. Cells were immunostained with γ-tubulin and counterstained with DAPI. Total anaphases were counted (50) and the proportion of multipolar anaphases is shown. Data are the mean ± SEM of six (Jurkat) and three (SR) independent experiments. (E) Jurkat cells were treated with 0.25 μM pevonedistat or vehicle control for 48 hours. Apoptosis was determined by Annexin-V staining. Data are mean ± SEM of 6 independent experiments. *p < 0.05 and **p < 0.01 vs. control.
Credit
Correspondence to - Alexey V. Danilov - adanilov@coh.org
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CC BY