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. 2001 Oct 15;155(2):271–278. doi: 10.1083/jcb.200107001

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Copyright © 2001, The Rockefeller University Press

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Figure 8. — Detection of PDE5 phosphorylation. (A) Purified unphosphorylated and phosphorylated GST–PDE5 fusion proteins (∼85 kD; 200 ng, each) were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were incubated with the antiphospho-PDE5 antibody in a dilution of 1:10,000. (B) Platelets were stimulated with 300 μM GSNO. At the indicated time points, aliquots (3.6 × 10⁷ platelets) were removed and proteins were separated by SDS-PAGE and transferred to nitrocellulose. Membranes were incubated with phospho-PDE5–specific antibody in a dilution of 1:50,000.