Figure 3.

LPS sensitizes fibroblasts to TGF-β1 stimulation of profibrotic gene expression. Fibroblasts were incubated with TGF-β1 (10 ng/mL) in the presence or absence of LPS for 24 hours. A: Total RNA was examined by qPCR. Results, normalized with GAPDH mRNA, are shown as the mean ± SD of triplicate determinations. B: Whole cell lysates and culture supernatants were subjected to Western blot analysis. Representative immunoblots are shown. C: Fibroblasts were immunostained with antibodies to type I collagen (Cgn I) or α-SMA or stained with DAPI, and viewed by immunofluorescence microscopy (top panels) or laser-scanning confocal microscopy (bottom panels). Original magnification, ×400. D: Right panel: Mean immunofluorescence intensity of individual fibroblasts stained with α-SMA was determined in five different microscopic fields. Results are given as the mean ± SD. *P < 0.05.