TLR3 expression in liver after APAP challenge.

<p>(<b>A</b>) Quantitative real-time PCR (TaqMan) was performed to measure the transcript levels of TLR3 in whole liver homogenates at 0, 4, 16, 24 and 48 h after the injection of APAP. Data shown indicate mean ± SEM of two independent experiments (<i>n</i> = 9–10 per time point). (<b>B</b>) Confocal immunofluorescent examination of livers revealed TLR3+ cells (red) and the keratin 8+ cells (green) surrounding the areas of necrosis in WT mice at 24 h after APAP injection. Shown are representative sections from 1 mouse of 5 per group. (<b>C</b>) The expression of TLR3 was analyzed in primary hepatocytes by western blotting. Each lane represents isolated hepatocytes from independent experiments. (<b>D</b>) Expression of TLR3 transcript levels in primary hepatocytes. 1.5×10⁵ cells/well were plated in 24 well-plates and stimulated with the indicated stimulus (5 mM of APAP, 20 ng/ml of TNFα, 10 µg/ml of PolyI:C) for 24 h. Bars represents mean±SEM of three independent experiments. *p<0.05 when APAP+TNFα+PolyI:C was compared with APAP+TNFα; ***p<0.0001 when only PolyI:C was compared with medium. Immunofluorescence analysis of livers revealed TLR3+ cells (red) in (<b>E</b>) Kupffer cells (F4/80), (<b>F</b>) CD34, and (<b>G</b>) CD11c in liver sections from WT mice at 24 h after APAP. Magnification: 400×. Shown are representative sections from 1 mouse of 5 per group.