Figure 3: The biological effects of the NO donor BNN6 on TILs. Note: (A) The effects of N, N’-di-sec-butyl-N,N’-dinitroso-1,4-phenylenediamine (BNN6) for the proliferation of tumor infiltrating lymphocytes (TILs) were measured by a Cell Counting Kit-8 (CCK-8) test. The results indicated weak cytotoxicity to TILs at concentrations below 33.3 μL/mL. (B) The enzyme-linked immunosorbent assay (ELISA) method for interferon-γ (IFN-γ) detection of TILs secretion (NC is negative control with 0 μL/mL BNN6). (C) Analyzed spots by inspection under an ELISPOT plate reader for IFN-γ detection of TILs secretion (the number shows the positive cells; C1, C2, C3, C4 were the 0, 3, 10, and 30 μg/mL BNN6 treatments, respectively). (D) Calculation of the IFN-γ secretion of 0, 3, 10, and 30 μg/mL BNN6 treated TILs using the ELISPOT method. Data are expressed as the mean ± SD. *P < 0.05, **P < 0.01 (paired t-test).