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. 2004 Nov;124(5):527–540. doi: 10.1085/jgp.200409153

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Copyright © 2004, The Rockefeller University Press

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Figure 6. — Inhibition of GC or PKG prevents ATP-dependent polypeptide phosphorylation and dephosphorylation in intact tissue. Epithelial tissue was preincubated for 10 min in the absence (lanes 1 and 2) or in the presence of either 50 μM LY-83583 (GC inhibitor, lanes 3 and 4) or 1 μM KT-5823 (PKG inhibitor, lanes 5 and 6) followed by stimulation with extracellular ATP (100 μM) for 5 min (lanes 2, 4, and 6). Cilia were then isolated and subjected to Western blot analysis, as described under materials and methods. Application of LY-83583 did not alter the basal level of phosphorylation (lane 3), although preincubation with KT-5823 resulted in a moderate phosphorylation of 44-kD polypeptide (lane 5), which was not influenced by the ATP addition (compare lanes 5 and 6). A representative of seven similar experiments is shown.