Figure 5.

The cross-talk between the TLR4 and Fn14 modulate TNF-α expression in LPS-stimulated SIEC02. Western blot analysis of TNF-α expression in the LPS-stimulated SIEC02 in the presence of increasing concentration of anti-Fn14 (A) or both anti-Fn14 and anti-TLR4 (B). The expression of TNF-α in SIEC02 treated with different concentrations of isotype antibody was used as control (C). Western blot analysis of the activity of MyD88 and NF-κB pathways in siRNA-mediated Fn14 gene knockdown SIEC02 stimulated with LPS in the absence (D) or in the presence of (E) TLR4 antibody. Representative blots are shown in the upper panel and the histograms in the under panel show the results of densitometric analysis of indicated gene expression (normalized to β-actin to correct for protein loading). The data represent the mean ± SEM (n = 3). ^*P < 0.05, ^**P < 0.01, compared with data on untreated cells. Super Fn14 expression on siRNA-mediated Fn14 gene knockdown SIEC02 stimulated with LPS in the absence or in the presence of TLR4 antibody was analyzed by flow cytometric analysis (F).