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. 2023 Sep 22;6(12):e202302156. doi: 10.26508/lsa.202302156

Figure S6. In vitro 4-OH tamoxifen-generated Themis-deficient CTL exhibit reduced cytolytic activity and cytokine production.

Figure S6.

The cytotoxic activity of Themis-deficient CTL generated using tamoxifen was assessed using a cell-based cytotoxicity assay. Antibody to CD107a was also added before stimulation to measure degranulation. (A) Specific lysis of peptide-loaded EL4 cells by tamoxifen-treated and untreated OT-I Themis cWT (right panel) and Themis cKO (left panel). (B) Surface expression of CD107a from the same samples in (A). Cytokine production was induced by stimulating CTL with EL4 cells loaded with different OVA APLs for 6 h before intracellular cytokine staining. (C) Intracellular IFNγ levels in tamoxifen-treated and untreated OT-I Themis cWT and cKO CTL. Percentage of IFNγ-positive cells. (D) Intracellular TNF expression of the same samples in (C). Percentage of TNF-positive cells. CTL were pooled from three mice per genotype. Data are representative of three experiments. Samples were compared using two-way ANOVA with multiple comparison test, with **** indicating P-value < 0.0001, *** P-values from 0.0001 to 0.001, ** values from 0.001 to 0.01, and * values from 0.01 to 0.05. Values shown here are mean ± SD.