Figure S4.

ACADVL overexpression confers enhanced metabolic fitness. Related to Fig. 6. (A) Transduction efficiency, gated on live CD8⁺ Thy1.1⁺. (B) Expression of the indicated activation receptors on transduced cells, gated on live CD8⁺ Thy1.1⁺ Vβ9⁺ cells. The gray histogram represents naive P14 staining, the red histogram represents TCR₁₀₄₅-transduced T cells, and the blue histogram represents ACADVL-TCR₁₀₄₅ transduced T cells. Data are representative of three independent experiments. (C) Production of IFNγ and TNFα by TCR₁₀₄₅ T (red) and ACADVL-TCR₁₀₄₅ T cells (blue) 7 d after activation and transduction, stimulated with Msln_406–414 peptide for 5 h. (D) Proportions of cotransferred TCR₁₀₄₅ T (red) and ACADVL-TCR₁₀₄₅ T cells (blue) before and after transfer (day 21) in tumor, blood, or spleen, gated on live CD8⁺ Thy1.1⁺ cells. Plots are representative of four independent experiments. (E) Inhibitory receptor expression on engineered cells isolated from tumor at day 10, gated on live CD8⁺ Thy1.1⁺ Vβ9⁺ cells. The gray histogram represents naive P14 staining, and the blue histogram represents ACADVL-TCR_1,045–transduced T cells. (F) Ex vivo production of IFNγ and TNFα by TCR₁₀₄₅ T (red) and ACADVL-TCR₁₀₄₅ T cells (blue) isolated from the spleen (SP) or tumor (tum) 10 d after transfer and stimulated for 5 h with Msln_406–414 peptide. Data are representative of four independent experiments. In all graphs, error bars represent mean ± SEM. Statistic was calculated using a two-tailed Student’s t test; ns, not significant.