Fig. 3.

Effect of H. hepaticus infection on expression and localization of Toll-like receptor-4 (TLR4) in the ileum. A: TLR4 mRNA levels evaluated using real-time PCR. The mean steady-state mRNA level for the FF group was assigned a value of 1.0, and mean mRNA levels for the FF+H.hep group were determined relative to this number. Values are means ± SE; n = 22–23 animals/experimental group. ^‡P < 0.02 vs. FF. B: representative TLR4 (95-kDa) bands from Western blot analyses are shown for FF (n = 7) and FF+H.hep (n = 7) groups. All samples were analyzed on the same gel. Densitometric values were normalized to β-actin and to FF value. *P ≤ 0.01 vs. FF. C: representative slides from FF and FF+H.hep groups stained for TLR4 (green) were evaluated using confocal laser scanning microscopy (n = 4 animals/experimental group). In the FF group, a weak TLR4 signal (green) was observed in cells in the lamina propria. In the FF+H.hep group, a strong TLR4 signal was localized mainly in cells in the lamina propria and some enterocytes. Cell nuclei were stained with YOYO-1 (blue). No signal was observed in negative control sections (data not shown). Magnification: ×400.