NMCMs were differentially stimulated and stained with LysoTracker Red DND-99 (300 nM, Thermo Fisher Scientific, L-7582) for 10 min according to the manufacturer’s protocol. The excess LysoTracker was removed with culture medium. The cells were fixed and permeabilized. Coverslips were mounted using ProLong Gold antifade reagent (Thermo Fisher Scientific). Images were captured using a fluorescence microscope at 40× magnification (Zeiss) and analyzed using Image-Pro Plus v6.0 software.
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