Meiosis-specific prophase-like pathway controls cleavage-independent release of cohesin by Wapl phosphorylation
Fig 3
Rec8 phosphorylation is required for efficient dissociation of Rec8 at late prophase-I.
(A) Chromatin fractionation assay for cdc20-mn (KSY642/643) and cdc20-mn cdc5-mn (KSY659/660) mutant cells was carried out as described in Fig 2A. (B) Quantification of Rec8 band intensity in (A) is performed as shown in Fig 2B. Error bars show the S.D. (n = 3). (C) Localization of Rec8 (red) and Rec8-pS521 (phospho-S521; green) was analyzed in cdc20-mn (KSY642/643) cdc20-mn cdc5-mn (KSY659/660) cells at 5, 8 and 12 h. (D) Total Rec8, Rec8-pS521, and DAPI signal intensity was studied as in Fig 1C. Error bars show the S.D. (n = 3). (E) Localization of Rec8 (red) with or without DAPI (blue) in CDC20-mn (KSY642/643) and CDC20-mn CDC5-mn (KSY659/660) mutants. Representative image is shown. The bar indicates 2 μm. (F) Kinetics of Rec8 in (E) was classified as shown in Fig 1B. Error bars show the S.D. (n = 3). (G) Total Rec8 and DAPI signal intensity was quantified as shown in Fig 1C. A minimum 30 nuclei were quantified in each representative time points. Error bars show the S.D. (n = 3).