The authors of “Multiple determinants and consequences of cohesion fatigue in mammalian cells” (Mol. Biol. Cell [2018] 29, 1811–1824; originally published in MBoC In Press as 10.1091/mbc.E18-05-0315) wish to make a correction to Figure 2. In the original HTML and PDF versions, the schematic is incorrect in Figure 2C. This has been corrected in the revised figure below.
FIGURE 2:
Wapl-mediated release of cohesin after metaphase is not required for cohesion fatigue. (A) Wapl depletion during interphase slows cohesion fatigue in subsequent mitosis. Elapsed times from metaphase to chromatid separation/cohesion fatigue were determined via live-cell imaging in HeLa cells stably expressing SMC1-GFP. In Wapl-depleted cells, only cells with clear GFP signal on chromosomes at the metaphase plate (indicating successful Wapl depletion) were scored in two independent experiments with totals of >100 cells. (B) Expression of phosphorylation-resistant sororin slows cohesion fatigue. Elapsed times from metaphase to cohesion fatigue were determined in HeLa cells expressing either wild-type sororin or nonphosphorylatable 9A-sororin. At least 60 cells were scored for each cell type. The Mann–Whitney test was used for statistical analysis. (C) Inhibition of Aurora B kinase after metaphase alignment does not inhibit cohesion fatigue. Experimental scheme and graph depicting elapsed times from metaphase to cohesion fatigue were determined after 2.5 μM ZM 447439 treatment in cells released from nocodazole to MG132 for 1 h. Three independent experiments with totals of >200 cells were quantified. The Mann–Whitney test was used for statistical analysis. (D) Locking the SMC3-Rad21 gate but not other gates before mitotic entry inhibits cohesion fatigue. Experimental scheme and results from chromosome spreads in Hek293 expressing cohesin fusions to rapamycin-binding proteins treated with rapamycin to lock specific gates before cells entered mitosis and then treated with MG132 for 6 h to arrest cells at metaphase and allow cohesion fatigue. Totals of >100 spreads per condition per cell line were quantified. Graph shows mean ± SEM. Dotted line represents the expected inhibition of fatigue with efficient SMC3-RAD21 gate locking based on the percentage of spreads with unresolved chromatid arms (45%) from Supplemental Figure 2D. (E) Locking any of the cohesin gates after completion of Wapl-mediated cohesin release in early mitosis does not inhibit cohesion fatigue. HEK293 cells expressing cohesin fusions to rapamycin-binding proteins were treated with or without rapamycin after allowing completion of early mitosis, Wapl-mediated cohesin removal in three independent experiments with totals of >450 spreads per cells line for each treatment. Graph shows mean ± SD. Two-way analysis of variance (ANOVA) with Tukey’s multiple comparison test was used for statistical analysis.
The HTML and PDF versions were corrected on the Molecular Biology of the Cell website on September 24, 2018. These corrections may not appear on copies of the article that reside on other websites.