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. 2013 Jan;182(1):192–205. doi: 10.1016/j.ajpath.2012.09.007

Figure 5.

Figure 5

LPS synergy with TGF-β1 is TLR4 dependent. A: Normal skin fibroblasts were incubated with LPS and TGF-β1 in the absence or presence of TLR4 inhibitors. Total RNA was examined by qPCR. B: Whole cell lysates and secreted proteins in the culture media were subjected to Western blot analysis. Type I collagen (Cgn I) protein levels were quantitated. Values shown indicate relative levels corrected for GAPDH in each lane. C: Normal skin fibroblasts were transfected with TLR4-specific siRNA or scrambled (Scr) control siRNA and incubated with LPS and TGF-β1 for 24 hours. Whole cell lysates were examined by using Western blot analysis. Representative immunoblots. Cgn I levels were quantitated. Values shown indicate relative levels corrected for tubulin in each lane. D and E: Skin fibroblasts from WT mice and TLR4-mutant (C3H/HeJ) mice were incubated in parallel with LPS and TGF-β1 for 24 hours. Whole cell lysates were subjected to Western blot analysis (D). Fibroblasts were fixed, incubated with antibodies to α-SMA, and examined by immunofluorescence microscopy (E). Blue, nuclei were identified by DAPI. Original magnification, ×100. *P < 0.05.