Imaging and Quantification of Germline

Images (0.2 μm sections) were acquired at 40×, 60× and 100× magnification of optically bisected germlines^{20 (link)} using a Deltavision, deconvolved, and merged using softWorRx (Applied Precision). Exposure conditions were kept constant for each strain and condition. Germlines regions were determined as described^{50 (link)} and the fluorescence intensity for each region was measured using ImageJ (version 2.0.0) and normalized to the number of nuclei in the given region. 3–8 germlines were quantified for each strain and condition. Student’s t test statistical analyses were performed using GraphPad Prism. Images were further processed using ImageJ and Adobe Photoshop CS6. In Fig. 2, 78–302 mitotic cells, 59–125 transition zone cells and 60–323 pachytene cells were quantified. Quantitative analysis of RAD-51 foci presented in Fig. S5 was performed as described^{51 (link)}.