JCS02466 Supplementary Material

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• Supplemental Figure 1 - Fig. S1. DNA-PKcs is not essential for phosphorylation of H2AX during prophase. Meiotic surface spreads of Prkdc^SCID/SCID spermatocytes stained for SCP3 and gH2AX are indistinguishable from the wild type.

• Supplemental Figure 2 - Fig. S2. Prophase-stage distribution analysis of the Atm^–/– Spo11^+/– mutant. Spermatocyte spreads from a 29 dpp Atm^–/– Spo11^+/– mouse and a wild-type littermate were stained for SCP3 and gH2AX. Over 500 nuclei were classified in the different stages of prophase I as leptonema (L), zygonema (Z), pachynema (P), diplonema (D) and diakinesis (Dk), according to axial-element morphology. The bar graph shows that the proportion of nuclei undergoing the different stages of prophase I in both genetic backgrounds is equivalent. We also analysed a 39 dpp Atm^–/– Spo11^+/– spread preparation and obtained comparable results.

• Supplemental Figure 3 - Fig. S3. In Spo11^–/– and Atm^–/– Spo11^–/– spermatocytes, BRCA1 localizes to the chromosomal cores immersed in gH2AX-chromatin domains. Surface-spread preparations from wild-type (a-f), Spo11^–/– (g-l) and Atm^–/– Spo11^–/– (m-o) spermatocytes were triple stained with antibodies against SCP3, BRCA1 and gH2AX. (j-l) Magnified regions of (g-i), respectively. BRCA1 stains the cores of gH2AX-chromatin domains in both mutants, suggesting that it might recruit ATR to these chromatin domains.