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Luminex cytokine profiling assay

CM Cody S. Madsen
AM Ashley V. Makela
EG Emily M. Greeson
JH Jonathan W. Hardy
CC Christopher H. Contag
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Cell culture supernatant was stored at −20 °C until ready for use. Supernatant was analyzed for CCL2 (MCP-1), CCL3 (MIP-1a), CXCL2/MIP-2, G-CSF, IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-12p40, IL-12p70, IL-13, TNF-α and VEGFα cytokine expression. Cytokine levels of cell supernatants were measured using a MCYTOMAG-70K-17 Mouse Cytokine Magnetic Multiplex Assay (Millipore Sigma) using a Luminex 200 analyzer instrument (Luminex Corp, USA) according to the manufacturer’s instructions. Standard one-way ANOVA with Tukey post-hoc test was used to determine statistically different values amongst all treatment groups.

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