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8944239513
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For in situ hybridization, brains from 6-day-old B6CBA mice were dissected free of the skull and immediately frozen. Sagittal sections were cut, transferred to coated slides, and fixed in paraformaldehyde. Sections were hybridized with digoxigenin-labeled RNA probes with the use of reagents from Boehringer Mannheim according to the manufacturer's protocol. After hybridization at 59°C, sections were washed and incubated with an alkaline phosphatase-labeled antibody to digoxigenin (Boehringer Mannheim) and developed in substrate buffer for 2 to 6 hours in a volume of ≃ 100 μl per slide. Each 100 μl of hybridization buffer contained 100 ng of probe and 50% formamide, 5x standard saline citrate, yeast RNA (1 mg/ml), 1x Denhardt's, 0.1% Tween-20, 0.1% CHAPS detergent, and 5 mM EDTA. Additional information about the expression of GIRK1 and GIRK2 is available at http:// peterson02.mc.duke.edu.
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8944221296
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note
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+] increased the number of cells transfected with wvGIRK2 or wvGIRK2-GIRK1 that survived. NMDG-containing media did not change the expression of wild-type GIRK2 or GIRK1.
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8944251708
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+, filtered at 5 kHz (8-pole Bessel filter), and digitized at 25 kHz (pCLAMP6).
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0028267224
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0026446347
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L. Heginbotham, Z. Lu, T. Abramson, R. MacKinnon, Biophys. J. 66, 1061 (1994); L. Heginbotham, T. Abramson, R. MacKinnon, Science 258, 1152 (1992). P. A. Slesinger et al. [Neuron 16, 321 (1996)] have reported a loss of selectivity of truncated wvGIRK2 and wvGIRK2-containing channels.
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0030045830
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L. Heginbotham, Z. Lu, T. Abramson, R. MacKinnon, Biophys. J. 66, 1061 (1994); L. Heginbotham, T. Abramson, R. MacKinnon, Science 258, 1152 (1992). P. A. Slesinger et al. [Neuron 16, 321 (1996)] have reported a loss of selectivity of truncated wvGIRK2 and wvGIRK2-containing channels.
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Slesinger, P.A.1
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8944262933
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βγ, and S. Manahan for assistance with cell culture. Supported by a Colciencias award to B.N., a National Research Service Award award to M.E.K., a Minnesota American Heart Association award to B.V., and an NIH grant to D.E.C. A.S.P. is a Sandoz scholar and is supported by a grant from the National Down Syndrome Society.
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