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Ca2+ levels were measured using RHOD-2AM (20 nM) at 548/574 nm (excitation/emission wavelengths); mitochondrial membrane potential (MMP) was investigated using TMRM (1 μM) at 552/581 nm and NADH autofluorescence was measured at 350/470 nm. Gates were applied using unstained controls. Cells were analysed on a BD LSRFortessa™ analyser (BD Biosciences, USA) using the BD FACSDiva software (BD Biosciences, USA).
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