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. 2018 Jun 27;9:1491. doi: 10.3389/fimmu.2018.01491

Figure 3.

Figure 3

Mutation on the cleavage site of toll-like receptor 9 (TLR9) decreases cleaved TLR9 and its response. (A) Amino acid sequence of WT or deletion mutant of TLR9 around cleaved sites. Triangles in the figure indicate the cleaved sites. The cleaved sites were analyzed by Edman degradation. (B) Cleavage pattern of TLR9 with mutation in the cleavage site. FLAG-6xHis tagged-WT or mutant TLRs were expressed in stem cell-derived DCs (scDCs) obtained from Tlr9−/− mice. FLAG tag was precipitated and C-terminal region of TLR9 was detected. The intensity of the bands was quantified and shown in the table. (C,D) scDCs expressing WT or mutant TLR9 were stimulated by TLR9 or TLR4 ligand. After 24 h incubation, IL-6 in culture medium was detected by ELISA. Cells were stimulated in triplicated wells and the mean SD is shown. At least two times of experiments were performed independently (B–D). Data were statistically analyzed by two-way ANOVA with multiple comparisons (C,D). ****p < 0.0001. Abbreviations: N/A, not applicable.