Figure 1. Characterization of the calcineurin regulatory subunit in WT and Ppp3r2 knockout mice.

A) Genotyping of knockout mice by PCR using primers designed to the detect the presence of knock out allele (containing LacZ reporter gene), as described in the materials and methods section, shows absence of Ppp3r2. Figure in the right panel is the western blot analysis of sperm cell extracts from KO and wild type mice using specific antibodies shows the absence of PPP3R2 in KO mice. B) Immunostaining of adult testis sections shows absence of PPP32 in knockout mice while it is present within the seminiferous tubules of the testis of wild type mice; green arrow indicates the absence of PPP3R2 in Sertoli cell. Hoechst dye was used to identify nuclei. C) Sperm permeabilized with 0.5% Triton X-100 and fixed with paraformaldehyde and stained with rabbit PPP3R2 antibodies followed by Alexa fluor-488 labelled secondary antibodies. The merged figures in the left of each panel shows localization of the protein mainly in the midpiece and dimly in the principal piece of sperm from wild type sperm while there is no detectable signal in sperm from knock out mice.