FIG. 4.

rsc2Δ mutant cells exhibit defects in establishment of sister chromatid cohesion. (A) Wild-type cells (Wt; RSC2) and rsc2Δ mutants expressing either Smc3-HA (YKB353 or YKB355) or Scc1-HA (YKB342 or YKB341) were blocked in G₁ by α-factor treatment and released at 37°C. Chromosome spreads shown are of cells were taken prior to release or at 100 min postrelease. DNA was stained with DAPI (4′,6′-diamidino-2-phenylindole). Chromatin-associated Scc1-HA and Smc3-HA were detected by indirect immunofluorescence. (B) Wild-type (YPH1477; circle), scc1-73 (YKB426; triangle), and rsc2Δ (YKB235; square) cells were arrested with α-factor and released into nocodazole media at 25°C. Cultures were shifted to 37°C after >90% of cells had budded (time zero), samples were taken every 20 min for 180 min, and the percentages of sister chromatid separation were scored. One hundred cells were scored per sample; two separate experiments were performed with nearly identical results.