Fig.2. scFv from TR1419 CAR mediated tumor cell apoptosis by binding with TRAIL-R1. (A) Schematic representation of TR1419∆ζ and the lentiviral vector. The construct consisted of the extracellular portion of anti-TRAIL-R1 scFv, linking via a CD8α hinge-transmembrane domains to the truncated CD3ζ molecule. (B) The expression of TR1419-28BBζ and TR1419∆ζ in Jurkat cells was detected by flow cytometry. (C) Cytotoxicity of TR1419-28BBζ- and TR1419∆ζ -Jurkat cells against SW480 in the absence or in the presence of soluble TRAIL-R1 protein was determined by RTCA. The analysis was performed using Students’ t tests. Data reflected the mean ± SEM of triplicate wells, ***p < 0.01. (D) SW480 were fixed and labeled with annexin V and 7-AAD and then analyzed by flow cytometry to identify apoptotic cells (annexin V+/7-AAD and annexin V+/7-AAD+), after incubation with effector cells for 2h. (E) The expression of caspase3 protein in SW480 analyzed by flow cytometry, after incubation with effector cells for 2h.