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Manual patch clamp recordings

DL Dominik Loser
MH Maria G. Hinojosa
JB Jonathan Blum
JS Jasmin Schaefer
MB Markus Brüll
YJ Ylva Johansson
IS Ilinca Suciu
KG Karin Grillberger
TD Timm Danker
CM Clemens Möller
IG Iain Gardner
GE Gerhard F. Ecker
SB Susanne H. Bennekou
AF Anna Forsby
UK Udo Kraushaar
ML Marcel Leist
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Manual patch clamp experiments were performed in the whole-cell mode (Hamill et al. 1981) with an EPC 10 USB patch clamp amplifier and PatchMaster Software (version 2 × 90.5; HEKA Elektronik, Lambrecht, Germany). Extracellular solution contained [mM]: 140 NaCl, 4 KCl, 1 MgCl2, 1.8 CaCl2, 10 HEPES and 10 d-glucose, pH 7.4. Intracellular solution contained [mM]: 107 K-gluconate, 10 KCl, 1 MgCl2, 10 HEPES, 5 EGTA, 4 Na2ATP and 0.2 NaGTP, pH 7.2. Recordings were executed at room temperature. For agonist tests in current-clamp and voltage-clamp mode, cells were kept at a holding potential of − 70 mV and the compounds were applied for 5 s.

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