Pe conjugated anti vimentin antibody clone d21h3

The cells were stained for the surface expression of CD34 using a BV421-conjugated anti-CD34 antibody (clone 581, BD Horizon, Franklin Lakes, NJ, USA), the expression of CD90 using an FITC-conjugated anti-CD90 antibody (clone 5E10, BioLegend, Koblenz, Germany), the expression of vimentin using a PE-conjugated anti-vimentin antibody (clone D21H3, Cell Signaling Technology, Frankfurt, Germany) and the expression of CXCR4 using a PE-conjugated anti-CXCR4 antibody (clone12G5, R&D Systems, Biotechne, Wiesbaden, Germany). The cells were also stained with an APC-conjugated OV-6 antibody (R&D Systems, Biotechne, Wiesbaden, Germany), Data were acquired on a BD FACSCanto II (BD Biosciences, San Jose, CA, USA) and analyzed using the Flowjo software v7.2.5 (BD Life Sciences, Ashland, OR, USA).

The HuH6 and HepG2 cells were stained for the expression of CD34 using a BV421-conjugated anti-CD34 antibody (clone 581, BD Horizon, Franklin Lakes, NJ, USA); the expression of CD90 using a FITC-conjugated anti-CD90 antibody (clone 5E10, BioLegend, Koblenz, Germany); the expression of Vimentin using a PE-conjugated anti-Vimentin antibody (clone D21H3, Cell Signaling Technology, Frankfurt, Germany) and the expression of PD-L1 using an APC-conjugated anti-PD-L1 antibody (clone 130021, R&D systems, Biotechne, Wiesbaden, Germany). The cells were also stained with an APC-conjugated OV-6 (Oval Cell marker) antibody (R&D Systems, Biotechne, Wiesbaden, Germany) or a PE-conjugated OV-6 antibody (Santa Cruz Biotechnology, Heidelberg, Germany). Flow cytometry analyses were performed using a BD FACSCanto II (BD Biosciences, San Jose, CA, USA). All data were analyzed using the Flowjo software v7.2.5 (BD Life Sciences, Ashland, OR, USA).