Fig. 5

The α-1,3 fucosylation of TNFR1 mediated by FUT10 impacted the binding capacity to TNF-α. a Chondrocytes were transfected with siRNA and treated with or without TNF-α for 48 h, and the protein level of TNFR1 was determined by Western blotting. β-Tubulin was used as an internal control. b The relative expression level of TNFR1 from three experimental replications was normalized to β-tubulin and compared based on one-way ANOVA. c Immunoblot and lectin blot reactivity of TNFR1 immunoprecipitated from chondrocytes treated as described above. d The fucosylation of TNFR1 was normalized to the expression of TNFR1 and compared using one-way ANOVA. e Immunoblot and lectin blot reactivity of TNFR1 immunoprecipitated from cartilage of OA patients and normal controls. f The fucosylation of TNFR1 was normalized to the expression of TNFR1 and compared based on a paired t test. g A schematic diagram of the fabrication of the TNFR1 antibody microarray. h TNFR1 was immunoprecipitated from chondrocytes transfected with siRNA (low level of α-1,3 fucosylation) or induced with TNF-α (high level of α-1,3 fucosylation), and the scanned images were obtained for the analysis of the binding ability of TNFR1 with different levels of fucosylation to TNF-α. The spots of the TNFR1 antibody are marked with white boxes. i The fluorescence intensities of spots were extracted by Genepix 7, and the binding ability of TNFR1 was compared based on fold change and t test. The data are presented as the mean ± SEM, *p < 0.05, **p < 0.01, and ***p < 0.001