Fig. 6. LY294002-induced tyrosine phosphorylation of AKAP3. (A,B) Western blot analysis of tail protein extracts from swim-up-selected spermatozoa treated or not with 10 µM LY294002, separated on 10% SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was first blotted with anti-AKAP3 antibody (A) and, after stripping, re-probed with anti-phosphotyrosine antibody, PY20-HRP (B). (C,D) Western blot analysis of total protein extracts from swim-up-selected spermatozoa treated or not with 10 µM LY294002. Twenty µg of sperm lysates were run in duplicate onto 8% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was cut longitudinally in two parts that were probed with anti-FSP95 (C) and anti-AKAP3 (D) respectively. Molecular mass markers are indicated on the right of the blots. Representative of three similar experiments.