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For live cell imaging, neutrophil-like differentiated cells were plated in 1- or 4-well glass-bottom chambers (Nalge Nunc International, Naperville, IL, USA), coated with 0.2% gelatin at 37°C for 1 h. Chambers were washed, and cells were plated. Cells were stimulated at the 1 µM concentration of fMLP. The difference in intracellular fluorescent proteins was directly imaged using a confocal microscope. Images were exported and analyzed with ZEN software (Zeiss, Jena, Germany).
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