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. 2017 Feb 10;31(10):2151–2160. doi: 10.1038/leu.2017.8

Figure 1.

Figure 1

Organization of the CD5CAR NK-92 construct and its expression. (a) Schematic representation of the CD5CAR NK-92 lentiviral vector. The CD5CAR NK-92 construct is a modularized signaling domain containing: a leader sequence, an anti-CD5scFv, a hinge domain (H), a transmembrane domain, two co-stimulatory domains (CD28 and 4-1BB) and the intracellular signaling domain CD3 zeta. (b) Western Blot analysis of CD5CAR. GFP (lane 1) and CD5CAR NK-92 (lane 2) were used to transduced HEK-293FT cells and subsequently probed with mouse anti-human CD3 zeta antibody 48 h post transduction for Western blot analysis. The expected weight of the CD5CAR NK-92 construct is 57.86 kDa by sequence analysis data (not shown). (c) Flow cytometry analysis of CD5CAR NK-92 expression on NK-92 cell surface (right) vs vector control NK-92 cells (left) and CD5CAR isotype control (middle). The population in green delineates transduced NK-92 cell population after fluorescence activated cell sorting. Gating was based off the vector and isotype controls.