Analysis of the Relationships between DNA Double-Strand Breaks, Synaptonemal Complex and Crossovers Using the Atfas1-4 Mutant
(A) Expression analysis of genes encoding meiotic proteins in WT and Atfas1-4 bud samples. Values are the average of assays carried out in triplicate using five different cDNA preparations. The red line is the reference for the fold change respect to the WT after normalization to 18S rRNA. The means corresponding to changes in gene expression and their standard errors are indicated. (B-Q) Dual immunolocalization of AtASY1 and AtZYP1 with HR proteins in WT and Atfas1-4 prophase I nuclei. (B, C) AtASY1 (green) and γH2AX (red) on WT and Atfas1-4 at leptotene. (E, G) AtASY1 (green) and AtRAD51 (red) on WT and (F, H) on Atfas1-4 at G2 or leptotene, respectively. (J, K) AtASY1 (green) and AtDMC1 (red) on WT and Atfas1-4 at leptotene. (M, N) AtASY1 (green) and AtMSH4 (red) on WT and Atfas1-4 at late zygotene. (P, Q) AtZYP1 (green) and AtMLH1 (red) on WT and Atfas1-4 at pachytene. (S, T) AtZYP1 (green) and AtMUS81 (red) on WT and Atfas1-4 at pachytene. Bars = 5 µm. (D, I, L, O, R, U) Total foci per nucleus in WT (red) and Atfas1-4 (blue) PMCs are indicated. Each dot is the count from a single nucleus. P values are from two-sided Wilcoxon Mann-Whitney tests (***P < 0.001).