The full-length, truncated TRPC5 constructs or empty vector was transfected into HEK293T cells (from the ATCC), together with an mCherry plasmid. Cells with red fluorescence were selected for whole-cell patch recordings (HEKA EPC 10 USB amplifier, PATCHMASTER 2.90 software). A 1-s ramp protocol from −100 to +100 mV was applied at a frequency of 0.2 Hz. Signals were sampled at 10 kHz and filtered at 3 kHz. The pipette solution contained 130 mM CsCl, 1 mM MgCl2, 5.7 mM CaCl2, 10 mM EGTA, and 10 mM Hepes (calculated free Ca2+, 200 nM), and the pH was titrated to 7.2 using CsOH. The standard bath solution contained 140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM Hepes, and 10 mM d-glucose, and the pH was adjusted to 7.4 with NaOH. The recording chamber (150 μl) was perfused at a rate of ~2 ml/min. All recordings were performed at room temperature.

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