Fig. 4: Mdivi-1 improves the cytotoxic function of CTLs targeting autologous cancer cells in vitro.

A Scheme of tumor-specific CTLs generation and in vitro coculture. B Twenty hours later, Th1 cytokines production was quantified by specific ELISAs (mean ± s.e.m; n = 5; p < 0.0001 for HNSCC and NSCLC; **p < 0.001 compared with non-coculture group by two-tailed t-test). C Residual E/T ratio was evaluated by flow cytometry after 3 days based on CD8 and EpCAM expression (mean ± s.e.m; n = 3; p = 0.0015 or p < 0.0001 for HNSCC and p < 0.0001, p < 0.0001 for NSCLC; *p < 0.01, **p < 0.001 compared with E/T ration = 1:1 group by two-tailed t-test). D Evaluation of MHC-I membrane expression in HNSCC and NSCLC primary cancer cells by flow cytometry. F indicates the fold change of MFI normalized to mock (mean ± s.e.m; n = 12; p = 0.0484, 0.0001, 0.0005 for HNSCC and 0.0487, 0.0001, 0.0001 for NSCLC ^#p < 0.05, *p < 0.01, **p < 0.001 compared with mock by two-tailed t-test). E Cell viability of primary cancer cells and PI uptake of tumor-specific CTLs after sequential treatment by MTS and flow cytometry. F Twelve hours after coculture, EpCAM⁺ cancer cells were harvested and cell death was examined with flow cytometry based on the uptake of PI and Annexin V. E/T ratio was 5:1 (mean ± s.e.m; n = 4; p < 0.0001 for HNSCC and p < 0.0001 for NSCLC; **p < 0.001 compared with mock by two-tailed t-test). G Th1 cytokines production was quantified by specific ELISAs after 24 h. E/T ratio was 5:1 (mean ± s.e.m; n = 5; p < 0.0001 for HNSCC and NSCLC; **p < 0.001 compared with mock by two-tailed t-test). H Evaluation of residual E/T ratio by flow cytometry based on CD8 and EpCAM expression respectively following 3 days. E/T ratio was 5:1 (mean ± s.e.m; n = 3; p < 0.0001, p = 0.0011, 0.0009 for HNSCC and p < 0.0001, p = 0.0015, 0.0012 for NSCLC; *p < 0.01, **p < 0.001 compared with mock by two-tailed t-test). E indicates effector cells, namely T cells. T indicates targeted cells, namely cancer cells.