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. Author manuscript; available in PMC: 2022 May 29.
Published in final edited form as: Nat Cancer. 2021 Nov 29;3(1):60–74. doi: 10.1038/s43018-021-00280-y

Fig. 2. MTDH inhibits T cell activation.

Fig. 2

a, Schematic diagram of in vitro tumor-immune cell co-culture assay. Ovalbumin (OVA) expressing Py8119 tumor cells were seeded into plates. One day after the seeding, splenocytes or CD8+ T cells isolated from OT-I mice were co-cultured with tumor cells at 10:1 ratio. 24 hr after co-culture, the cells or culture media were collected for the following analysis. b,c, Indicated tumor cells were co-cultured with CD8+ T cells. 24 hr after co-culture, culture media were collected for cytotoxicity analysis (b). The tumor cells were harvested for western blot to examine with apoptotic markers PARP and cleaved-caspase 3 (CC-3) (c). Data represent mean ± SEM. n=3 independent experiments. Significance determined by one-way ANOVA analysis with Sidak’s test for multiple comparisons. d-f, OT-I splenocytes co-cultured with indicated tumor cells for 24 hr were harvested for flow cytometry analysis. The expression of activation markers CD137 (d) and IFN-γ (e) in CD8+ T cells were determined. MFI, Mean Fluorescence Intensity. The culture media were collected for ELISA assay to examine the concentration of IFN-γ (f). Data represent mean ± SEM. n=3 independent experiments. Significance determined by one-way ANOVA analysis with Sidak’s test for multiple comparisons. Numerical source data for b, d-f, and uncropped blots for c are provided.