Extended Data Fig. 7: PSA for IgE isotype controls and characterization of NEUFcε.
From: Sialylation of immunoglobulin E is a determinant of allergic pathogenicity
a, Temperature change following OVA-induced PSA in mice receiving DNP-specific SiamIgE on day 0, and then PBS, OVA-specific SiamIgE or OVA-specific AsmIgE isotype controls (from Fig. 3e) on day 1. n = 4 mice for all groups; two-way ANOVA with Tukey’s multiple comparison test. b, Protein gel stain (left) and immunoblot (IB) for mIgE (right) of native and denatured NEUFcε. c, Binding kinetics of analyte NEUFcε to ligand hFcεRIα on biosensor. Kinetics of analytes of threefold serial dilution from 26.2 nM to 0.32 nM were analysed. d, FACS analysis of surface-bound NEUFcε on LAD2 mast cells following 30 min of sensitization at 37 °C. n = 3 technical replicates per group, representative of two independent experiments. e–h, Neuraminidase activity of NEUFcε determined by digestion of mIgE or fetuin overnight (e–g), and detection of protein loading by Coomassie blue (e), terminal α2,6-sialic acid by SNA (f), and terminal galactose by Erythrina cristagalli lectin (ECL) (g) or by the amount of substrate 2-O-(p-nitrophenyl)-α-d-N-acetylneuraminic acid digested by NEUFcε in a colorimetric assay (h; n = 3 technical replicates per group, representative of two independent experiments). Data are means ± s.e.m.
