Extended Data Fig. 4: RSC facilitates overexpressed Rec8 localization to meiotic centromeres, and expression of Spo11 and Rec8 in proliferating cells causes KT loss as visualized via a fluorescent array on cen1.
From: Centromeres are dismantled by foundational meiotic proteins Spo11 and Rec8
a, Frames of representative films monitoring overexpressed Rec8-GFP through meiosis. During prophase, Rec8 localizes throughout the nucleus. As cells approach MI, Rec8 begins to disappear but remains concentrated at centromeres. At the end of MI, Rec8 foci are clearly visible in the wt cell, but not in rsc1Δ cells. The frames highlighted in the yellow box are enlarged in the right-hand panels; these foci subsequently disappear at MII. Scale bars, 5 μm. b, Quantification of meiotic cells harbouring Rec8 foci at the end of MI. Total numbers of films analysed are indicated above each bar. All rsc1Δ films show diminished Rec8-GFP at the end of MI, and most fail to show any detectable Rec8-GFP on at least one SPB. c, Top: schematic outlining the loss of KT components on cen1 in anaphase cells. Below: images of live cells in late anaphase; the KT is visualized with Ndc80-2xGFP, cen1 via cen1-tetO/R-tomato, and the spindle via mCherry-Atb2. Arrows indicate lagging cen1 with KT loss. Lagging cen1 with intact KT is also shown (right-hand panel) but is excluded from quantification. Scale bars, 5 μm. d, Quantification of cen1 KT loss as exemplified in a. Expression of Spo11 or Rec8 increase the anaphase KT loss of cen1. Total numbers of cells analysed are indicated above each bar. P values are determined by two-tailed Fisher’s exact tests and indicated above brackets.
