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. 2018 Jun 6;13(6):e0198347. doi: 10.1371/journal.pone.0198347

Fig 3. Specific cytotoxicity of FOLR1-CAR KHYG-1 cells against FOLR1-positive cells.

Fig 3

KHYG-1 cells were infected with lentivirus encoding the FOLR1-CAR or mock vector and were selected with puromycin for 2 weeks. (A) FOLR1-CAR expression was measured by western blot using the CD3ζ antibody in KHYG-1 cells. (B) The cytotoxic effects of KHYG-1 cells on K562 cells transfected with FOLR1 or mock vector were measured by luciferase assay at an E/T ratio of 10:1 for 4 h. (C) The lysis percentages of mock KHYG-1 and FOLR1-CAR KHYG-1 cells against KB cells were determined in a dose-dependent manner for 4 h using the luciferase assay. (D−E) To determine the lysis percentages of FOLR1-CAR KHYG-1 cells against GC cells, mock KHYG-1 and FOLR1-CAR KHYG-1 cells were co-cultured with (D) FOLR1-positive GC cells or (E) FOLR1-negative GC cells at an indicated E/T ratio for 4 h. Experiments were repeated at least three times with similar results. The data are represented as the mean rate of lysis ± SD (%) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired t-test (***P<0.001).