FIGURE 2.
Acute CCl4 induces ER stress and TBK1-mediated phosphorylation of IRF3 in hepatocytes. WT mice received a single injection of corn oil or CCl4. A, at the indicated time points, mice were sacrificed; splicing of Xbp1 mRNA (sXBP1) in the liver, indicating ER stress, was evaluated by PCR; and products were separated on 3% agarose gel. B, 2 h after CCl4 administration, hepatocytes (Hep.), or LMNCs from WT mice were isolated and analyzed for sXBP1 by PCR. C and D, at the indicated time points, phosphorylation of IRF3 was assessed by immunoblotting in liver (C), whereas Ifnb1 mRNA expression in liver and as serum IFN-β were measured by RT-PCR and ELISA, respectively (D). n = 4 mice per time point. E, primary hepatocytes isolated from WT mice were treated with 1 μm thapsigargin and were pretreated with DMSO or BX795 (10 or 100 μm), and analyzed for phospho-IRF3.