Cells complete karyo- and cytokinesis after a <i>cdc14-1</i> release, irrespective of the cXIIr segregation status.

<p>(A) Strain FM518 (<i>cdc14-1 tetO:487 TetR-YFP</i>) was first arrested at 37°C for 3 hours and micrographed in the conditions described to see the tetR-YFP nucleoplasm bridge (0′, see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002509#pgen.1002509.s005" target="_blank">Figure S5</a>). Then it was released at 25°C. Part of the yeast culture was released into fresh medium containing alpha-factor to arrest the daughter cells in G1. At 90 minutes after the release, more photos were taken and representative cells are shown. The two main cell morphologies at that time for a normal release, dumbbells [D] and foursomes [F], are depicted. The hollow triangle points to the nucleoplasm bridge. White pointers indicate the missegregated <i>tetO</i>. Note that cells have no nucleoplasm bridge. (B) Time course of nucleoplasm bridge disappearance for the same strain relative to the cXIIr segregation status. (C) Strain FM515 (<i>cdc14-1 RAD52-YFP</i>) was arrested in telophase by incubation at 37°C for 3 hours. Then the cell culture was shifted back to 25°C to enter a new cell cycle. At the time of the telophase block and 2 hours after the release, samples were taken and fixed with formaldehyde. Contrasted bright field micrographs of representative cells before and after zymolyase treatment are shown. Left corner photos show cells at the telophase block. The white triangle highlights the difference in bud neck thickness after zymolyase treatment. Main photos depict cells 2 hours after the telophase release. “F” points to foursomes, “D” to dumbbells and “S” to single cells. No foursomes (<3%) were seen after zymolyase treatment. (D) Strain FM515 (<i>cdc14-1 RAD52-YFP</i>) was treated to inhibit cytokinesis (+LatA) and/or cell cycle progression beyond G1 (+αF). The number of cells was counted in a haemocytometer after the zymolyase treatment. The chart represents cell number two hours after the release relative to their telophase block (mean ± SEM, n = 3).