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Fig. 4. Dose- and time-dependent effects of PI 3-kinase inhibition on tyrosine phosphorylation of sperm p110 kDa protein. Western blot analysis of sperm lysates from swim-up-selected spermatozoa stimulated for 1 hour with increasing concentrations of LY294002 (LY, A), or treated with 10 µM LY294002 for the different times indicated (B) were separated by 8% SDS-PAGE. Equal amounts (30 µg) of sperm proteins were subjected to 8% SDS-PAGE. Tyrosine phosphorylated proteins were revealed with PY20-HRP antibody. C, control at 10 minutes. Molecular mass markers are indicated on the right of the blots. Arrowheads indicate the modulated band at about 110 kDa. Representative of three similar experiments.





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