Figure 2
CD23 and DC-SIGN are functionally expressed on transfected HEK Freestyle cells, but do not bind human IgG1, regardless of sialylation status. (a–c) HEK Freestyle cells were transfected with human FcγRIIa (CD32, red), CD23 (blue), DC-SIGN (green), or an empty vector (Mock, pink) and tested for binding to anti-CD32, anti-CD23. or anti-DC-SIGN, or their natural ligands at 37 °C: asialylated IgG1, IgE, or ICAM-3, respectively. (d) Binding of different soluble IgG1 glycoforms (10 μg/ml) to HEK CD23, DC-SIGN, FcγRIIa and Mock transfected cells at 37 °C. Data are representative of three independent experiments (in duplicate) and shown as mean values of the geometric mean fluorescence intensity (gMFI ± s.e.m.) with the background signals subtracted. Significant differences compared to HEK Mock signals were determined by One-way ANOVA with Dunnett’s multiple comparisons test (*P < 0.05, **P < 0.01, ****P < 0.0001).
