Figure 3: Tenascin-C-induced fibrotic responses are TLR4-dependent.

(a–c) Human foreskin fibroblasts were incubated in media with tenascin-C (TNC; 2 μg ml⁻¹) in the absence or presence of CLI-095 or MyD88 blocking peptide or scrambled controls for 72 h. (a,c,d) Whole-cell lysates analysed by western blotting. Representative immunoblots. S, secreted; L, lysates. Band intensities, normalized for tubulin, shown below. (b–d) Immunofluorescence (IF) microscopy using antibodies to αSMA, and DAPI. Representative images; scale bar, 50 μm. One way analysis of variance followed by Bonferroni’s multiple comparison test. (d) Skin fibroblasts isolated from mice with tamoxifen-inducible fibroblast-specific TLR4 knockout (TLR4^−/−) and control mice (that is, no tamoxifen) incubated in media with tenascin-C (2 μg ml⁻¹) for 72 h. Upper panel, representative immunoblots. Lower panels, immunofluorescence; scale bar, 25 μm. Relative fluorescence intensities represent means from four randomly selected h.p.f.’s. Con, control.