MED12 mutation activates Notch signaling to enhance cancer stemness and suppress CD8+ T cell cytotoxicity in lung cancer

Figure 7 Mediator complex subunit 12 mutation mediated the inhibitory effect of cancer cells on CD8+ T cell killing and chemotaxis through activation of the Notch pathway. After co-culturing CD8+ T cells with NCI-H322 and PC9 cells that had been treated with or without DAPT, the following parameters of the CD8+ T cells or tumor cells were analyzed. A: Transwell assay was performed to assess the migratory capacity of CD8+ T cells toward tumor cells; B: The expression of interferon-γ, tumor necrosis factor-α, perforin, and granzyme B in CD8+ T cells was measured by enzyme-linked immunosorbent assay; C: Flow cytometry was used to detect the percentage of programmed cell death 1-positive CD8+ T cells; D: Flow cytometry was used to detect the percentage of programmed death ligand-1-positive NCI-H322 and PC9 cells; E: The level of lactate dehydrogenase (LDH) in tumor cells was measured by LDH Cytotoxicity Assay Kit to evaluate the cytotoxic activity of CD8+ T cells. Data are presented as mean ± SD. All experiments were repeated 3 times. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. NS: Not significant; IFN: Interferon; TNF: Tumor necrosis factor.