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elucidate the cause of meiotic arrest, we analyzed meiotic chromosomes with a variety of markers that are diagnostic of recombination and synapsis. Recombination in Trip13Gt/Gt spermatocytes appeared to initiate normally as judged by the presence of γH2AX in leptonema (Figure S2A and S2B), which reflects the presence of meiotically induced DSBs [18]. RAD51 and/or DMC1, components of early recombination nodules (ERNs), was also present as abundant foci in Trip13Gt/Gt zygotene spermatocytes (unpublished data; the anti-RAD51 antibody cross-reacts with DMC1), indicating that recombinational repair of DSBs is initiated. The cohesin complex, which is essential for completion and/or maintenance of synaptic associations, appeared to assemble normally as judged by immunolabeling for the meiosis-specific cohesins STAG3 (Figure S2C and S2D) and REC8 (unpublished data). Because yeast PCH2 localizes to telomeres in a Sir3p-dependent manner, we tested for possible telomere defects by immunolabeling for TRF2, a component of a protein complex that plays an essential role in telomere protection [19]. It was localized to telomeres of both fully synapsed and telomerically asynaptic mutant chromosomes (Figure S2E and S2F)."}

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{"project":"2_test","denotations":[{"id":"17696610-11242108-85497900","span":{"begin":351,"end":353},"obj":"11242108"},{"id":"17696610-9476899-85497901","span":{"begin":1099,"end":1101},"obj":"9476899"},{"id":"T44728","span":{"begin":351,"end":353},"obj":"11242108"},{"id":"T98903","span":{"begin":1099,"end":1101},"obj":"9476899"}],"text":"To elucidate the cause of meiotic arrest, we analyzed meiotic chromosomes with a variety of markers that are diagnostic of recombination and synapsis. Recombination in Trip13Gt/Gt spermatocytes appeared to initiate normally as judged by the presence of γH2AX in leptonema (Figure S2A and S2B), which reflects the presence of meiotically induced DSBs [18]. RAD51 and/or DMC1, components of early recombination nodules (ERNs), was also present as abundant foci in Trip13Gt/Gt zygotene spermatocytes (unpublished data; the anti-RAD51 antibody cross-reacts with DMC1), indicating that recombinational repair of DSBs is initiated. The cohesin complex, which is essential for completion and/or maintenance of synaptic associations, appeared to assemble normally as judged by immunolabeling for the meiosis-specific cohesins STAG3 (Figure S2C and S2D) and REC8 (unpublished data). Because yeast PCH2 localizes to telomeres in a Sir3p-dependent manner, we tested for possible telomere defects by immunolabeling for TRF2, a component of a protein complex that plays an essential role in telomere protection [19]. It was localized to telomeres of both fully synapsed and telomerically asynaptic mutant chromosomes (Figure S2E and S2F)."}

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Recombination in Trip13Gt/Gt spermatocytes appeared to initiate normally as judged by the presence of γH2AX in leptonema (Figure S2A and S2B), which reflects the presence of meiotically induced DSBs [18]. RAD51 and/or DMC1, components of early recombination nodules (ERNs), was also present as abundant foci in Trip13Gt/Gt zygotene spermatocytes (unpublished data; the anti-RAD51 antibody cross-reacts with DMC1), indicating that recombinational repair of DSBs is initiated. The cohesin complex, which is essential for completion and/or maintenance of synaptic associations, appeared to assemble normally as judged by immunolabeling for the meiosis-specific cohesins STAG3 (Figure S2C and S2D) and REC8 (unpublished data). Because yeast PCH2 localizes to telomeres in a Sir3p-dependent manner, we tested for possible telomere defects by immunolabeling for TRF2, a component of a protein complex that plays an essential role in telomere protection [19]. It was localized to telomeres of both fully synapsed and telomerically asynaptic mutant chromosomes (Figure S2E and S2F)."}