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. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Methods Cell Biol. 2018 May 1;144:185–231. doi: 10.1016/bs.mcb.2018.03.008

Figure 9. Analysis of spindle assembly checkpoint (SAC) signaling.

Figure 9.

(A) Schematics of two-cell C. elegans embryos either untreated or depleted of ZYG-1, a component required for centriole duplication. ZYG-1 depletion causes the generation of a monopolar spindle where the kinetochores facing away from the centrosome are devoid of attachment. (B) Representative image sequences of two cell embryos (AB cell) depleted of ZYG-1 in the presence of a temperature-sensitive APC/C allele (mat-3(or344)) at the permissive temperature. This treatment causes cells to arrest in a manner that is dependent on the essential SAC component MAD-3. The addition of the (mat-3(or344)) allele compromises APC/C activity and extends the duration of the mitotic arrest, allowing the detection of subtle perturbations (Bezler/Gonczy 2010, Kim 2017. (C) Quantification of mitotic duration from the experiment in (B). These perturbations are used as a sensitized assay for the SAC (Adapted from Kim et al. 2017).