Forty-one cytokines and chemokines were analyzed simultaneously with bead-based multiplex assay using Milliplex Map Human Cytokine/Chemokine Magnetic Bead Panel (HCYTMAG-60K-PX41, Millipore) on the Luminex FlexMap 3D instrument according to the manufacturer’s instruction. Briefly, cells were seeded in a T25 flask and media were refreshed when the cells approached 70% confluence. Cell culture supernatant was harvested 48 hours after media change and multiplex cytokine assay was performed on the Luminex FlexMap 3D instrument. The results were normalized to cell numbers in each flask.

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