Fig. 1

Characterization of genetically engineered PAR1CAR-T cells. A Schematic of a lentiviral vector (LV) expressing a chimeric antigen receptor (CAR). B Anti-PAR1 expression levels in human T cells transduced with LV particles were analyzed using flow cytometry by detecting expression of an αPAR1 antibody. Transduction efficiencies are plotted in histograms. The 84.5% transduction rate is not an average but an individual result that served as a minimum requirement in our laboratory protocol. C A sandwich ELISA was performed to evaluate the binding ability of the PAR1CAR-T PAR1 antigen using PAR1CAR- and mock-transduced 293 T cells. Non-transduced 293 T cells were employed as a negative control (blank; ** p < 0.01). D PAR1CAR-T cell expression percentages are plotted as bars, and cell fold-changes are expressed in line plots, compared to mock-transduced T cells and non-transduced CD3 T cells. Data were derived from experiments involving three independent healthy donors during a 2-week culture period to stimulate the recombinant human PAR1-His.Tag protein