Figure 5

Effect of the absence of N-glycans on the interaction of MP84 with host cells. (A) Scheme of the engineered MP84 protein lacking all four predicted N-glycosylation sites (NΔMP84). All asparagine (Asn) residues in N-glycosylation sites were exchanged with alanine (Ala) residues. (B) Expression analysis of MP84 lacking N-glycosylation sites (NΔMP84). The MP84 secreted by the WT and alg3Δ strains, and the MP84 lacking N-glycosylation sites were separated via SDS-PAGE and subjected to Coomassie staining, Western blot analysis using anti-His antibody, and lectin-blotting (GNA-AP). The raw data of SDS-PAGE, western blot analysis, and lectin-blotting in (B) were presented in Supplementary Information as Fig. S6. (C) Comparison of adhesion activities in host cells between MP84 and NΔMP84. Significant differences were obtained via one-way ANOVA (**P ≤ 0.01, ****P ≤ 0.0001). (D) Comparison of the secreted cytokines between BMDCs co-incubated with MP84 and NΔMP84. Significant differences were obtained via one-way ANOVA (**P ≤ 0.01).