Human macrophages were loaded with 1.25 μm of Fluo3-AM (Thermo Fisher Scientific) for 30 min at room temperature in loading buffer containing 150 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 20 mM Hepes, 10 mM glucose, 1× PowerLoad (Thermo Fisher Scientific) to solubilize the Fluo3-AM dye, and 2.5 mM probenecid (Thermo Fisher Scientific) for dye retention. Fluo3-AM was then removed, and the cells were chased in loading buffer without PowerLoad and probenecid for 30 min at room temperature. Cytosolic Ca2+ was monitored by flow cytometry (FACSCanto II) or with a Nikon A1 confocal microscope.

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