Measurement of cytokines and chemokines in the supernatant
Soluble mediators were assessed using the Inflammation panel from the OLINK Proteomics platform(14) in the supernatant (n = 4) at 0 hours (resting state) and at 3, 6 and 24 hours after activation with high and low-affinity IgE. Samples only containing cell culture medium were analyzed as a blank control. The normalized protein expression values used are calculated from Ct values of a proximity extension assay that has been shown to have superior sensitivity compared to other detection methods (15). Sensitivity and range of detection for each analyte is available on OLINKs home page.