Mouse anti human e cadherin

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).

Cells were plated onto glass coverslips, fixed with 4% paraformaldehyde for 10 min and permeabilized with 0.1% Triton X-100 in PBS for 15 min. Blocking solution was applied for 1 h at room temperature. Primary antibodies, mouse anti-human E-cadherin (Cell Signaling Technology), and rabbit anti-human vimentin (Cell Signaling Technology), were applied at 4 °C overnight. FITC-conjugated goat anti-rabbit and Cy3-conjugated goat anti-mouse secondary antibodies were loaded and incubated for 1 h at room temperature. Immunostaining signals and DAPI-stained nuclei were visualized at room temperature using a confocal microscope (FV10i; Olympus) equipped with a 10 × /0.30 NA objective lens (Olympus) and FluoView software (version 4.3; Olympus).