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. 2019 Aug 19;8:e47646. doi: 10.7554/eLife.47646

Figure 6. Proteasome and APC/C inhibition at anaphase onset induces an anaphase arrest in human hTERT-RPE1 cells.

Figure 6.

(a) Control hTERT-RPE1 cell expressing endogenous Cyclin B1-Venus and co-stained with SiR-DNA to visualize mitotic chromosomes. (b) MG132 addition just before or at anaphase onset (time 00:00), caused an anaphase arrest with detectable Cyclin B1 levels and condensed chromosomes. The arrest was sustained up to 6 hr (time window of acquisition). (c) pro-TAME addition at anaphase onset (time 00:00) showed no visible effect during anaphase. (d) Apcin addition at anaphase onset (time 00:00) induced a slight delay in DNA decondensation. (e) APC/C inhibition with a cocktail of pro-TAME and Apcin at anaphase onset (time 00:00) caused a strong anaphase delay (during the time window of acquisition – 5 hr). Cyclin B1 localization is highlighted with LUT ‘fire’. Scale bars are 5 μm. Time is in h:min. (f) Quantification of Cyclin B1-Venus (normalized to anaphase onset) in control (n = 7 cells), MG132 (n = 4 cells), pro-TAME (n = 7 cells), Apcin (n = 6 cells) and pro-TAME+Apcin (n = 4 cells).