FIG. 3.

Freshly isolated hepatocytes do not express TLRs. Primary hepatocytes and intrahepatic lymphocytes were isolated from HBV transgenic mice as previously described (16, 25) and stained with antibodies specific for TLR2, -3, and -9 (eBioscience) in combination with anti-CD11c (BD Pharmingen). Surface staining was performed to detect TLR2 expression, while intracellular staining was applied for TLR3 and -9 expressions (18). Cells were acquired using a FACSCalibur flow cytometer (Becton Dickinson), and data were analyzed using CELLQuest software (Becton Dickinson). Histograms represent TLR staining (gray) or isotype control staining (white) of hepatocytes (left panel). Corresponding TLR expression on CD11c+ cells of intrahepatic lymphocytes were shown as positive controls (right panel).