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Western Blot Analysis A total of 6 mice at 6 mo of age (wild type, n = 3; FORKO, n = 3) were used for Western blot analysis. One testis of each mouse was extracted and cut into four equal pieces. Each piece was then frozen in liquid nitrogen for subsequent protein extraction. The frozen pieces of testis were homogenized, suspended in lysis buffer containing detergent and a protease inhibitor cocktail, and the solution was centrifuged at 11 000 X g for 15 min at 4°C. The supernatant was removed and quantified for protein using the Bradford Assay (Bio-Rad Laboratories Inc., Richmond, CA). Equivalent amounts of solubilized protein (30 |xg) in SDS sample loading buffer were boiled for 5 min and electrophoresed on a 10% SDS polyacrylamide gel. Subsequently, the proteins were transferred onto nitrocellulose membranes for immunoblotting. Following incubation with the anti-ABP antibody at 1:500 (v/v) dilution and secondary antibody conjugated to horseradish peroxidase (1:1000 v/v), bands were revealed by chemiluminescence using the Pierce SuperSignal Western Blotting Kit (Pierce, Rockford, IL). Molecular weight markers were used to estimate the mass of the detected bands. The experiment was repeated a minimum of three times in duplicate. Equal protein loading was confirmed by Coom-assie staining. Quantitative analysis of Western blot was performed using Chemi-Imager 4.0 software (B&L Systems, Maarssen, The Netherlands), which measured the spot density of each protein band on the exposed film. Raw density data was subject to an unpaired sample /-test; P values < 0.05 were considered significant.