Cells were plated in a 96-well plate and grown to confluency. The protocol broadly followed previously published studies (84). Briefly, the medium was changed to a potassium buffer [125 mM KCl, 2 mM K2HPO4, 1 mM MgCl2, 20 mM Hepes, 5 mM glutamate, and 5 mM malate (pH 7.0)] with 0.1 μM Calcium Green-5N. After baseline measurements, digitonin (0.02%) was added to permeabilize cells, followed by the serial addition of 200 nM CaCl2 solution in the same potassium buffer. Measurements were conducted on a BioTek spectrophotometer, and experiments were conducted in quadruplicate. Data were collected using Gen 5 software.

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