Nuclear accumulation of Pch2 is linked to increased association of Pch2 with the SC and assemblies of SC components in ZIP1+ cells

Abstract

(A-B) Immunofluorescence of spread meiotic chromosomes at pachytene stained with anti-GFP antibodies (to detect GFP-Pch2; green), anti-Zip1 antibodies (red) and DAPI (blue). Representative nuclei are shown. In both, (A) and (B), cultures were mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction. In (B), Auxin (500μM) was also added 12 h after meiotic induction to degrade Orc1. Spreads were prepared at 19 h. Arrows point to the rDNA region (nlo) and Polycomplex (PC). Scale bar, 2 μm. The strain in (A) is: DP1717 (crm1-T539C GFP-PCH2). The strain in (B) is: DP1885 (orc1-3mAID crm1-T539C GFP-PCH2). (C) Percentage of nuclei containing polycomplexes in the experiments shown in (A and B). Between 10 to 20 nuclei were counted for each strain and condition. (D) Quantification of the different patterns of Pch2 localization in the experiment presented in Fig 3A and 3B. Representative cells are shown.