(A, B) Localization of SPATA22 (green) and SYCP3 (red) in wild type
(left) and Spata22Gt/Gt (right) spermatocytes (A)
and oocytes (B).(C, D) Localization of MEIOB (green) and SYCP3
(red) in wild type (left) and Spata22Gt/Gt (right)
spermatocytes (C) and oocytes (D). (E, F) Localization of RPA
(green) and SYCP3 (red) in wild type (left) and
Spata22Gt/Gt (right) spermatocytes (E) and
oocytes (F). Whereas SPATA22 and MEIOB are absent in mutant meiocytes, RPA
localization remains unaffected. (G) Immunoblotting analysis of
protein extracts prepared from D15 wild type (+/+), heterozygous (+/−),
and homozygous mutant (−/−) testes, as well as D15 B6 control
testes, with antibodies against MEIOB, RPA1, SYCP1 and DMC1. Expression of GAPDH
(bottom panel) served as a loading control to normalize protein expression
across genotypes. The molecular weight of each protein (in kDa) is indicated to
the left of the corresponding immunoblot. (H) Analysis of RNA
extracts prepared from D15 wild type (+/+), heterozygous (+/−), and
homozygous mutant (−/−) testes. Transcripts for all meiotic genes
assessed were amplified by RT-PCR. The name of the gene evaluated is indicated
to the left of each corresponding panel. Expression of Actb was
used to normalize the expression of each gene across genotypes. H2O,
water control. D, postnatal day.