(A) Representative images of Rec8 staining. Scale bar, 4 μm. (B to E) Quantification of the numbers of Rec8 lines (B), the total length of Rec8 lines per cell and corrected length with unpaired homologs (numbers in parentheses) (C), the average length of each Rec8 line per cell (D), and Pds5 fluorescence intensity (E) in WT and mutants. The numbers of nuclei measured from left to right, n = 105, 103, 104, 102, and 101, separately (B to E). pCLB2-PDS5-AID with 2 or 10 mM IAA compared with 0 mM IAA, P < 0.0001 (B, C, and E), P > 0.05 (D), t test. (F and G) Illustration of the tetO/TetR-GFP (F) and lacO/LacI-GFP (G) assays. (H) Representative images of the assay for homolog pairing and sister cohesion in the ndt80Δ background. A single GFP spot and two GFP spots indicate homologs are paired and unpaired, respectively. Sister cohesion defects are reflected by three or four GFP spots. (I and J) Quantification of homolog pairing and sister cohesion detected by tetO/TetR-GFP (H; n = 200, 203, 202, 205, and 200, separately) or LacO/LacI-GFP assay (I; n = 204, 200, 205, 201, and 201, separately). (K to N) Quantification of the number of Rec8 lines per cell (K), the corrected (in parentheses) and uncorrected total length of Rec8 lines per cell (L), the average length of each Rec8 line in individual cells (M), and the axis length of chromosome II (N), in cells with paired (one GFP focus) and unpaired (two GFP foci) chromosome II. Sample size, n = 58 (one GFP focus) and 68 (two GFP foci) (K to N). Error bar, means ± SD (B to E and K to N). P < 0.0001, t test (K to N).