Fig. 7. Western blot analysis of immunoprecipitated AKAP3 from spermatozoa treated or not with 10 µM LY294002. After 1-hour treatment, sperm samples were extracted in SDS buffer (see Materials and Methods). Cells lysates were immunoprecipitated (IP) using an anti-AKAP3 antibody, electrophoresed on 8% SDS, and immunoblotted with anti-phosphotyrosine antibody (IB: PY20-HRP, upper panels), stripped and re-probed with anti-AKAP3 antibody (IB: AKAP3, A, lower panel) or anti-FSP95 antibody (IB: FSP95, B, lower panel). Molecular mass markers are indicated on the right of the blots. Representative of three similar experiments.