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A
Schematic representation of the conditional alleles (loxfrt or lox) and the null allele (−) obtained upon Cre‐mediated recombination for Pds5A (top) and Pds5B (bottom). The position of the primers used for genotyping is indicated (blue arrows).
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B
Example of genotyping PCRs for Pds5A (top) and Pds5B (bottom) alleles using DNA obtained from testis of the indicated mice as well as DNA from mouse embryo fibroblasts as control. cKO mice carry the conditional allele(s) in homozygosis and a Cre‐ERT2 transgene and have been treated with TX to promote the translocations of the Cre recombinase to the nucleus. In most cKO mice, excision of the targeted exon is not complete and a weak band corresponding to the conditional allele can be still be detected in addition to the stronger band of the null allele. The sizes of the different PCR products are as follows: Pds5A wild‐type (+) 872 bp, loxfrt 778 bp and null (−) 414 bp; for Pds5B wild‐type (+) 706 bp, lox 859 bp and null (−) 415 bp.
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C
Immunoblot analysis of total extracts prepared from testes of mice of the indicated genotypes. Decreasing amounts of extract from testes obtained from wild‐type mice (WT) were loaded for comparison. Overall, elimination of Pds5B is less efficient than elimination of Pds5A both in the single and in the double cKO mice.
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D–M
Double immunolabeling of PDS5A (green, D–H) or PDS5B (green, I–M) and SYCP3 (red) in Pds5A cKO spread spermatocytes at the indicated prophase I stages (D–G and I–L) and in metaphase I bivalents (H and M).
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N–W
As in (D–M), but in Pds5B cKO spread spermatocytes at the indicated prophase I stages (N–Q and S–V), and in metaphase I bivalents (R and W).
Data information: In both cases, from mid‐pachytene onwards PDS5 proteins are not depleted. Sex bivalents (XY) are indicated. Scale bar, 10 μm.
