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Nevertheless, in the light of the obtained P-values and the number of statistical testes performed, we cannot exclude that our positive findings may be due to chance.
Laboratory testes performed to rule out pituitary or parathyroid tumors were serum fasting prolactin, ACTH, cortisol, growth hormone and insulin like growth factor1, T4, TSH, intact PTH, calcium and phosphorus.
Heterozygous and homozygous variant genotype carriers were combined for the analyses of interactions due to power-considerations. Therefore, in the light of the obtained P-values and the number of statistical testes performed, we cannot exclude that our positive findings are due to chance.
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Also, histopathological examinations of the testes were performed.
Scanning of both testes is performed in sagittal and transverse planes with size measurements.
Using a probe designed from NAT cloned from testes we performed Northern blot analysis of total RNA, which revealed that the level of NAT mRNA was higher in midgut, ovary and female accessory glands than in fat body and brain.
Hypertonic perfusions of testes were performed essentially as described [32].
When multiple testes were performed, however, only values below 0.0001 were considered significant.
Collection of embryonic testes was performed following institutional approval for appropriate care and use of laboratory animals, according to published protocols (Pastor et al., 2014).
Dissections of testes were performed as described above on three batches of Y M and III M males each (i.e., three biological replicates of each strain).
For testes, we performed immunohistochemical assays for anti-Müllerian hormone (AMH), as previously described (Lambrot et al. 2006a, 2006b), using an anti-AMH polyclonal antibody (provided by N. Di Clemente).
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