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. 2021 Mar 12;7(11):eabe7920. doi: 10.1126/sciadv.abe7920

Fig. 6. Pds5 regulates recombination frequencies but does not impair the recombination process per se.

Fig. 6

(A to C) Alterations in Pds5 protein levels alter RPA focus number. (A) Photos from the time point with maximal RPA focus number in each strain. Scale bar, 2 μm. (B) Quantification of RPA foci. (C) Maximal RPA focus number. Bar, SE (n = 3). SPM, sporulation medium. (D) Map of HIS4LEU2 hotspot locus. (E) Southern blot images for DSB assay at HIS4LEU2 locus. The asterisk, a possible DSB fragment from an additional DSB site. (F) Quantification of DSBs at 10 hours in SPM (DSBs accumulated to maximum levels in rad50S). % DNA is the percentage of total hybridizing DNA signal. (G) Zip3 focus number is highly correlated with axis length (data from Figs. 2 and 3). (H) Average CO number. Strains: pCUP1-PDS5/” with 0 μM (#1), 10 μM (#4), and 25 μM (#6) copper; PDS5/pCUP1-PDS5 with 0 μM (#2), 1 μM (#3), 5 μM (#5), and 10 μM (#7) copper; WT (#8). Bar, SD. n = 8, 6, 5, 7, 5, 7, 4, and 10 tetrads for strains #1 to #8, respectively. (I) γ values obtained from best-fit gamma distributions. Bar, 95% confidence interval. Strains and sample sizes are as in (H). (J) CoC analysis of CO interference. Data are from (H). (K) Frequencies of chromosomes absence of Zip3 focus. Strains: pCLB2-PDS5/pCUP1-PDS5 with 0.02 μM (#1), 0.05 μM (#3), 1 μM (#4), and 2 μM (#9) CuSO4; pCUP1-PDS5/” with 0 μM (#2), 0.5 μM (#6), 1 μM (#7), and 2 μM (#10) CuSO4; #5, #8, and 11# are PDS5/pCUP1-PDS5 with 0 μM (#5), 0.5 μM (#8), and 1 μM (#11) CuSO4; WT (#12). n = 100, 123, 112, 104, 98, 108, 95, 117, 104, 119, 103, and 115 nuclei, respectively. Bar, SE (n = 3). Compared with WT, P > 0.05, two-proportions test.