HEK293 cells were seeded onto poly-l-lysine–coated 25-mm glass coverslips. Cells were transfected using 9 equivalents of homemade polyethylenimine with mouse pcDNA3.1-YFP-TRPM7 or control pEGFP plasmids. Recordings were done 24 to 72 hours after transfection. Cells were held at 0 mV, and ramps from −100 to +100 mV (400 ms) were applied at a frequency of 0.2 Hz. Ramp data were acquired at 10 kHz and low-pass–filtered at 1 kHz. Experiments were performed at room temperature (22° to 26°C). Whole-cell TRPM7 cationic currents were measured using pipettes (1.6 to 2 megohms) filled with a solution containing 140 mM CsCl, 10 mM Hepes, 1 mM EGTA, 4 mM Na2ATP, and 0.3 mM Na3GTP (pH 7.2 to 7.3 and 295 to 300 mosmol/liter). The external solution, which was also used for calcium experiments in HEK293 cells, contained 110 mM NaCl, 5 mM KCl, 1.8 mM CaCl2, 0.5 mM MgCl2, 10 mM Hepes, and 5 mM glucose (pH 7.3 to 7.4 and 305 to 310 mosmol/liter).

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