Ab113112

Rabbit anti-human PD-L1 monoclonal antibody (ab205921), rabbit anti-human IFIT2 polyclonal antibody (ab113112), rabbit anti-human GAPDH monoclonal antibody (ab205921), mouse anti-human STAT1 monoclonal antibody (ab3987) and mouse anti-human STAT1 (phospho Y701) monoclonal antibody (ab29045) were purchased from Abcam (Cambridge, MA, USA). Mouse anti-human E-cadherin (Cell Signaling, Danvers, MA, USA), rabbit anti-human N-cadherin and rabbit anti-human Zeb1 (Santa Cruz, Dallas, TX, USA), goat anti-mouse IgG and goat anti-rabbit IgG (Sigma, St. Louis, MO, USA) were used for Western blotting analysis. Mouse anti-human E-cadherin (MAB-0589) and Vimentin (MAB-0178) monoclonal antibodies used for the immunohistochemistry assay were purchased from Maixin Biotechnology (Fuzhou, China). The HRP-labeled goat anti mouse/rabbit secondary antibodies (K500711) were purchased from Dako (Glostrup, Denmark). The RNeasy Mini Kit was purchased from Qiagen (Valencia, CA, USA), and SYBR Green Master Mix kits were purchased from TaKaRa (Dalian, China). AG490 (S1143, Selleck) were purchased from Selleck (Shanghai, China). RPMI-1640 medium and fetal bovine serum (FBS) were purchased from Gibco (Cambrex, MD, USA).

Infected tissues were lysed with RIPA buffer at 2 dpi. 15μg of lysed proteins were loaded on a 10% SDS-PAGE gel and then transferred onto PVDF membrane (Bio-Rad). The membranes were first blocked with 5% skim milk (AppliChem) in TTBS (10 mM Tris HCl, pH 7.5, 500 mM NaCl, 0.05% Tween 20) at RT for 30min and then incubated with primary Ab overnight at 4°C. The membranes were then washed thrice with TTBS and incubated with secondary antibodies conjugated to HRP at RT for 1h. The membranes were washed and later developed with Western Bright Sirius ECL system (Advansta Inc.) for 2 min. Immunoblot images were acquired using Fujifilm LAS 4000 luminescence imager. Primary antibodies used in Western blot include anti-Stat1 (612233, BD), -MDA5 (ENZ-ABS299-0100, Enzo Life Sciences), -MX1 (ab95926, Abcam), -IFIT2 (ab113112, Abcam) and -actin (MAB1501, Merck Millipore). All primary antibodies were diluted 1:1000 except MX1 diluted in 1:2000. Goat anti-rabbit (7074, Cell Signaling Technology) and horse anti-mouse (7076, Cell Signaling Technology) secondary antibodies conjugated to HRP were diluted in 1:5000 and used for chemiluminescence development.