On the basis of previous literature, we assumed that the data such as Fig. 3 (C, right panel, and D, right panel) have a normal distribution, and the data such as Figs. 4D, 5B (right panel), and fig. S7C (right panel) have a non-normal distribution. Two-sided Student’s t test was used for Fig. 3C (right panel). Mann-Whitney U test was used for non-normal distribution data such as Figs. 4D, 5B (right panel), and fig. S7C (right panel). One-sample t test was used for normalized data by specified column such as Fig. 3D (right panel). Statistical tests were also indicated in individual figure legends and supplementary figure legends.

Figure 3C (right panel). For RAD51, n = 203, 191, and 201 (at 0 hour) and n = 149, 130, and 105 (at 3 hours) cells were analyzed in each independent experiment. For rad51-2A, n = 166, 160, and 189 (at 0 hour) and n = 158, 126, and 125 (at 3 hours) cells were analyzed in each independent experiment. The data represent the means ± SD of triplicate experiments.

Figure 3D (right panel). Extracts from three independent experiments were analyzed for each strain. The data represent the means ± SD of triplicate experiments. P values were determined by the one-sample t test. Immunoprecipitated MAT locus DNA in Rad51 strain column was used as a hypothetical mean.

Figure 4D. For the RAD52 strain, a total of 200 cells were analyzed. For the rad52-T412A strain, a total of 200 cells were analyzed. For the rad52-T412E strain, a total of 200 cells were analyzed. Box plot was normalized by the median of Rad52 cell with whiskers from the 5th to the 95th percentile. P values were determined by the Mann-Whitney U test.

Figure 5B (right panel). For the RAD52 strain, a total of 207 cells were analyzed. For the rad52-T412A strain, a total of 204 cells were analyzed. Box plot was normalized by the median of the RAD52 strain with whiskers from the 5th to the 95th percentile. P values were determined by the Mann-Whitney U test.

Figure S7C (right panel). For the RAD52 strain, a total of 218 cells were analyzed. For the rad52-T412A strain, a total of 207 cells were analyzed. Box plot was normalized by the median of the RAD52 strain with whiskers from the 5th to the 95th percentile. P values were determined by the Mann-Whitney U test.

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