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. 2023 Aug 18;4(3):102522. doi: 10.1016/j.xpro.2023.102522

Figure 1.

Figure 1

G1 release assay

(A) Schematic of G1 release assay (MGY2150). The G1 release assay monitors the timing of anaphase onset when tensionless and unattached kinetochores are induced by Taxol treatment during spindle assembly. Cells are first synchronized in G1 using alpha-factor before being split into two cultures. Upon release from alpha-factor arrest one is treated with Taxol in DMSO and the other with only DMSO as a control. Samples are withdrawn and fixed over the subsequent 240 min (0, 60, 90, 120, 150, 165, 180, 195, 210, 240 min) and used to monitor anaphase onset via DAPI staining of DNA (blue spots). Note that if alpha-factor is included at 90 min post release from G1 the anaphase re-bud cells will be schmooing. If alpha-factor is omitted they will be small-budded.

(B) Results from the G1 release assay where symbols represent the mean ± SEM from 3 different experiments. Percent anaphase cells at each time point is calculated by (number of anaphase large budded cells + anaphase re-bud cells) / the total number of cells scored in the time point ∗ 100. For each timepoint 100–200 cells were scored per experiment.