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IMMUCHEM Double Antibody Corticosterone RIA kit (ICN, Irvine, CA) was used by the veterinary diagnostic lab at Cornell University. Corticosterone was extracted from ground adrenals with 0.5 ml of ethyl acetate at room temperature and spun at 11,000 rpm for 5 min, and the upper phase was saved, vacuum dried, and resuspended in 400 μl of resuspension buffer with 5% ethanol.
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note
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Plasmid constructs for cRNA expression in Xenopus oocytes were devised by modification of a mouse Slo (mSlo) expression construct (5). Rat adrenal RT-PCR products corresponding to ZERO, STREX-1, and STREX-2 variants were made with primers rSlo3 and a downstream primer with ah internal Xho I site (sequence: 5′-ACCGCTCGAGCTCTGGCGTGGCTCCTCCTG-3′) and cloned into unique Hind III and Xho I sites in the mSlo vector. Restriction analysis and sequencing were used to confirm constructs. mSlo differs from rSlo outside the rat insert at only 4 of 716 residues. Rasmids were linearized with Sal I and transcribed with Ambion T3 mMESSAGE kit.
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We thank R. Harris-Warrick, D. Baro, R. Knapp, A. Bass, and R. Hoy for helpful discussions. Supported by the American Heart Association
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We thank R. Harris-Warrick, D. Baro, R. Knapp, A. Bass, and R. Hoy for helpful discussions. Supported by the American Heart Association.
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