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Figure 5. Sildenafil down-regulates IL-4R expression on CD11b+ cells. CD4+, CD8+, CD11b+, or CD11c+ cells were magnetically purified from spleens of C26GM-bearing mice challenged 9 d earlier. CD4+ or CD8+ T cells were stimulated with ConA for 3 d, with or without 50 µg/ml sildenafil. (A) Purified cells were cultured separately for 2 d with or without sildenafil, harvested, and used to determine cGMP levels, as described in Materials and methods. (B) T cell proliferation was determined by [3H]thymidine incorporation. (C) CD11c+ cells were analyzed for B7.2 and MHC class II expression. (D) CD11b+ cells were analyzed for IL-4R expression by flow cytometry. The t test p-value is reported. Error bar values are shown. MFI, multiplicity of infection.
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Footnotes ?1 Abbreviations: PDE, phosphodiesterase; cAMP, adenosine 3?5?-cyclic monophosphate; cGMP, guanosine 3?5?-cyclic monophosphate; GAF, cyclic GMP, adenylyl cyclase, FhlA; CaM, calmodulin; PK, protein kinase; PKA, cAMP-dependent protein kinase; PKG, cGMP-dependent protein kinase; CNS, central nervous system; ANP, atrial natriuretic peptide; NO, nitric oxide; PDP, 9-(6-phenyl-2-oxohex-3-yl)-2-(3,4-dimethoxybenzyl)-purin-6-one; BAY 60-7750, 2-(3,4-dimethoxybenzyl)-7-[(1R)-1-hydroxyethyl]-4-phenylbutyl]-5-methylimidazo[5,1-f] [1,2,4]triazin-4(3H)-one; PI3K, phosphatidylinositol 3-kinase; IGF1, insulin-like growth factor 1; OPC-33450, 6-[3-[3-cyclooctyl-3-[(1R*,2R*)-2-hydroxycyclohexyl]ureido]-propoxy]-2(1H)-quinolinone; UCR, upstream conserved region; CRE, cAMP-responsive element-binding protein; RACK, receptor for activated C-kinase 1; AKAP, A-kinase anchoring proteins; ERK, extracellular signal-regulated kinase; BRL 50481, 3-(N,N-dimethylsulfonamido)-4-methyl-nitrobenzene; PAS, Period, Arnt, and Sim; IBMX, 3-isobutyl-1-methylxanthine; BAY 73-6691, 1-(2-chlorophenyl)-6-[(2R)-3,3,3-trifluoro-2-methylpropyl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4-one.
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