Osteoclast acid production was detected using Acridine orange (AO) stains as previously described 31 (link). Osteoclasts were incubated in a-MEM containing 5 mg/ml of Acridine orange (Sigma) for 15 min at 37 °C, washed, and chased for 10min in fresh media without Acridine orange in the dark. We use fluorescence microscope (Leica Texas Red filter) to observe stained cell.
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