Figure 6.
SIC interacts with TLR2 and CD14 to activate THP1 cells
(A) THP1 cells were pre-incubated for 30 min with IgG1 and Fc fragment and then incubated with 5 μg/mL SIC +/− 2.5% plasma (P). NF-κB activation was analyzed at 655 nm.
(B) Cells were pre-incubated with a neutralizing antibody against TLR2 and then incubated with 5 μg/mL SIC +/− 2.5% plasma (P). Activation of NF-κB was measured at 655 nm.
(C) Quantification of phosphorylation of Western blots of THP1 cells treated with a neutralizing antibody against TLR2. The band intensities of p-p38 samples were normalized to loading control (p38 band) and values analyzed.
(D) Quantitative DIA-MS analysis of THP1 cells before and after trypsin treatment.
(E) THP1 cells were pre-incubated with antibodies against CD14 and then incubated with 5 μg/mL SIC +/− 2.5% plasma (P). NF-κB activation was analyzed.
(F) Surface plasmon resonance data show interaction of different SIC concentrations to immobilized TLR2. Representative original data are displayed in colors, 1:1 Langmuir fitted curves are presented in black.
(G) Surface plasmon resonance data show interaction of different SIC concentrations to immobilized CD14. Representative original data are displayed in colors, 1:1 Langmuir fitted curves are presented in black. All data represent mean ± SEM of 3 independent experiments, one-way ANOVA, Dunnett's multiple comparison test, with single pooled variance. ∗∗p < 0.01, ∗∗∗∗p < 0.0001.