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. 2010 Jul 20;30(18):4391–4403. doi: 10.1128/MCB.00002-10

FIG. 5.

FIG. 5.

SPO11β is expressed in excess such that a reduction by 50% in its level of expression does not translate into a comparable decline in the number of meiotic DSBs. (A) SPO11α and -β polypeptides are reduced by one-half in Spo11+/− testes, irrespective of ATM deficiency. Total testis extracts from Spo11+/+ and Spo11+/− littermates as well as Atm−/− Spo11+/−, Atm−/− Spo11+/+, and Mlh1−/− mice were immunoprecipitated/blotted with anti-SPO11 antibody. * and ** (also for panel B), minor splicing isoforms of SPO11α. (B) Amounts of SPO11 isoform precipitated from mutant and wild-type extracts, taking into consideration the difference in total protein between extracts. For each isoform, the band intensity (in arbitrary units) was divided by the amount of total protein in the extract. The normalized values for the α and β isoforms are documented in the table. (C) Amount of SPO11 isoform precipitated from each extract normalized to the wild-type (wt) extract. For each isoform, the band intensity (in arbitrary units) was divided by the amount of total protein in the extract and this ratio was divided by the corresponding ratio obtained for the wt sample: (intensity/mg total protein)mutant/(intensity/mg total protein)wt. (D) A 50% reduction in SPO11β polypeptide does not translate into a comparable decline in the number of DSBs. Spermatocyte spreads from Spo11+/+ and Spo11+/− (12-dpp) pups were stained for SCP3 and RAD51. The graph reports RAD51 focus numbers from 22 and 19 nuclei, respectively.