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For Airyscan microscopy, cells in Nunc™ Lab-Tek™ 8-well chamber slides were washed once with pre-warmed phenol red-free DMEM (37 °C), then directly incubated with 50 nM TMRE in phenol red-free DMEM for 30 min at 37 °C in the dark (no post-staining wash to preserve ΔΨm and enable immediate imaging)