Figure 6

Western blot analysis of the five proteins. (A) HEK cells transfected with empty vectors (lane 1) or GFP-tagged cDNA sequences of the five genes (lane 2) were subjected to Western blot analysis, using antibodies to the proteins. The expected molecular weights of the proteins are 53 kDa (Mm.333010), 75 kDa (Mm.23534), 50 kDa (Mm.23377), 52 kDa (Mm.425335), and 57 kDa (Mm.116803). An anti-tubulin antibody (α-Tubulin) was used as a control antibody. (B) Protein samples from different stages (testicular spermatogenic cells, testicular sperm, and mature sperm) were blotted with antibodies. Two proteins (Mm.23534 and Mm.23377) displayed alterations in molecular weight. Among the five proteins, three present in sperm were analyzed with regard to their localization. Mature sperm were treated with biotin (+) or left untreated (-), and subjected to Western blot analysis. ADAM2 was used as a control protein. TC, testicular spermatogenic cells; TS, testicular sperm; S, mature sperm; ADAM, a disintegrin and metalloprotease.