Figure 2.
Analysis of Pcs1 localization. (A) Cells expressing Pcs1-GFP and Mis6-HA (JG15031) were permeabilized by zymolyase digestion, extracted with detergent, fixed and stained with antibodies against HA and GFP. Nuclei were visualized by Hoechst staining. (B) Untagged cells (JG12013) or cells carrying Pcs1-GFP (JG14985) or Sgo2-GFP (JG12239) were harvested at the indicated time-points (0 and 8 minutes) after release from nda3-KM311-arrest. Chromatin binding of the GFP-tagged proteins was analyzed by chromatin-immunoprecipitation followed by quantitative PCR using oligonucleotide primers specific for the centromeric central region (cnt2), outer centromere (dgII) and chromosome arm (top1).