Isotype control (IgG-PE) was purchased from BD pharmingen (BD Biosciences San Jose, CA, USA). Anti-JAK, anti-p53, anti-p21waf, and anti-survivin www.selleckchem.com/products/Bortezomib.html were purchased from Cell Signaling Technologies (Beverly, MA). Anti-beta-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). The STAT3 inhibitor WP1066 and the JAK inhibitor pyridone 6 were purchased from Merck (Darmstadt, Germany). Neutralising anti-EGFR and anti-IL-6R antibodies were purchased from Millipore (Billerica, MA) and R&D Systems (Minneapolis, MN) respectively. Recombinant glycoprotein gB was purchased from Abcam (Cambridge, UK). Ganciclovir was purchased from Roche (Basel, Switzerland). Cell culture HepG2 cells were obtained from the European Collection of Cell Cultures (ECACC, Porton Down, UK) and PHH from Kaly-Cell (Strasbourg, France).

HepG2 cells were cultivated in Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% fetal bovine serum, 1% non-essential amino acids (Sigma-Aldrich, St. Louis, MO), penicillin (100 IU/ml), and streptomycin (100 microg/ml). PHH were cultivated in serum-free Dulbecco’s Modified Eagle Medium supplemented with L-glutamine (2 mmol/l), insulin (4 microg/ml), dexamethasone (1 micromol/l), and gentamycin (50 mg/l). The PHH were free of HCV, HBV, HIV, and HCMV as determined by highly sensitive PCR and RT-PCR assays (Abbott; Argene). Cell viability assay was performed as previously described [21]. IL-6 production was measured in culture supernatants using an ELISA kit (Human IL-6 Quantikine ELISA kit, R&D Systems, Minneapolis, MN).

Quantification of HCMV titers in cell culture supernatants was performed by real-time PCR as previously described [11]. HCMV infection of HepG2 cells and primary human hepatocytes Cell-free virus stock was prepared by propagating two strains of HCMV, the laboratory strain AD169 and a clinical isolate, HCMV-DB, in MRC5 human fibroblasts as described previously [11], [21]. AD169 is a highly-passaged laboratory strain of HCMV originally isolated from the adenoids of a child [22]. The clinical isolate HCMV-DB was isolated from a cervical swab specimen from a 30-year-old pregnant woman [21]. MRC5 human fibroblasts were cultured in EMEM with 10% FBS, penicillin (100 IU/ml), and streptomycin (100 microg/ml). HepG2 cells and PHH were infected at different multiplicities of infection (MOI) for 2 h at 37��C, washed thoroughly, and covered with fresh medium.

Where specified, cells were treated with ganciclovir (5 microg/ml) during infection with HCMV. Ultraviolet (UV)-inactivated HCMV (UV-HCMV; 1200 microJ.cm?2, 15 min) was used as control. Supernatants were clarified by centrifugation and stored at ?80��C Cilengitide until use. Virus titers were determined by plaque-forming assay in MRC5 human fibroblasts as described previously [23]. RT-PCR assay Briefly, total RNA was extracted from HepG2 cells with RNeasy mini kit (Qiagen, Hilden, Germany).