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Fig. 1. Expression profile of Nanos proteins. (A) Characterization of
Nanos2 and Nanos3 antibodies. Cell extracts were prepared from E14.5 and E12.5
male gonads, resolved by SDS-PAGE and subjected to western blot analysis with
Nanos2 and Nanos3 antibodies, respectively. Arrows indicate the endogenous
proteins. (B) Expression profile of Nanos proteins in embryonic gonads.
Extracts were prepared from each single male embryonic gonad of wild-type and
Nanos2-/- littermates. One-half of these extracts were resolved by
SDS-PAGE and subjected to western blot analysis with Nanos2 and Nanos3
antibodies. Actin was used as a reference for protein amounts. (C)
Analysis of Nanos mRNA and protein expression. Total RNA and protein extracts
were prepared from E14.5 male gonads of wild-type and Nanos2-/-
littermates. Expression of Nanos2 and Nanos3 mRNA was
analyzed by RT-PCR. G3PDH was used as the internal standard for normalization.
The expression levels of the Nanos2 and Nanos3 proteins were analyzed by
western blotting as in B.
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