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3H]thymidine was then added per well. Sixteen hours later, cells were harvested for scintillation counting.
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note
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The three PDE7 antisense oligonucleotides were as follows: from position 1 to 24 (AS-0: 5′-CGGCAGCT-GCTAACACACTTCCAT); from position 708 to 728 (AS-708: 5′-CAGTGCATGGCCTGAGTAAC); and from position 937 to 959 (AS-959: 5′-GGCAGATGT-GAGAATAAGCCTG). For RT-PCR analysis, PDE7-specific primer pairs were as follows: 5′-GATATTTGTA-ACCCATGTCGGACG-3′ and 5′-AAAGCTTGGCGG-TACTCTATCGAT-3′. PDE4A-specific primer pairs were as follows: AAGAGGAGGAAGAAATATCAATGG and TTACAGCAACCACGAATTCCTCC.
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We thank L. H. Li for help with RT-PCR analysis and D. Stenger for comments on the manuscript. This research is supported by NIH grant DK21723 to J.A.B. and by a grant from the Ono Pharmaceutical Company. C.Y. is the recipient of a Career Award from the Burroughs Wellcome Fund.
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