Figure 3.

Workflow for NK-92 transduction, selection, and cloning

NK-92 were first transduced with the HER2 synNotch lentiviral vector and sorted twice for constitutive HER2 synNotch expression. Subsequently, cells were transduced with the GAL4-driven CEA-CAR lentiviral vector and sorted to exclude those with basal CEA-CAR expression after co-culture with HER2-normal CRC cells. Finally, cells were co-cultured with HER2amp cells and sorted for positive CEA-CAR expression, to concomitantly generate a sorted population and individual clones. (A) Transduction and sorting workflow. (B) Flow cytometry plots before and after each sorting step, as indicated. Red boxes represent the selected populations. y axis, log10 expression; x axis, forward scatter; %, percent of cell above the depicted positivity threshold; MFI, mean fluorescence intensity.