Extended Data Fig. 6: Cellular and biochemical characterizations of wildtype and mutant γδ TCR–CD3 complexes.

a, The expression level of γδ TCR or CD3 on Jurkat-76 cells expressing Vγ9Vδ2 TCR (Cαβ/CPαβ), Vγ9Vδ2 TCR (Cγ1), and Vγ9Vδ2 TCR (Cγ2). b, The expression level of γδ TCR or CD3 on Jurkat-76 cells expressing Vγ5Vδ1 TCR (Cγ1), and Vγ5Vδ1 TCR (Cγ2). c, Quantitative detection of cholesterol sulfate by LC–MS/MS. The characteristic peaks of cholesterol sulfate in the Vγ9Vδ2 TCR–CD3 complex sample and the standard sample, denoted by numerals. Specifically, the peak at m/z 465.30 corresponding to parent ion [M + H-H2O]⁺. d, Quantitative detection of cholesterol by LC–MS/MS. The characteristic peaks of cholesterol in the Vγ9Vδ2 TCR–CD3 complex sample and the standard sample, denoted by numerals. Specifically, the peak at m/z 369.43 corresponding to parent ion [M + H-H2O]⁺. e, The representative chromatogram of cholesterol sulfate (up) and cholesterol (down) extracted from a purified Vγ9Vδ2 TCR–CD3 complex sample. The abundance of cholesterol is nearly 70-fold higher than that of cholesterol sulfate in purified Vγ9Vδ2 TCR–CD3 proteins. f, The expression level of γδ TCR or CD3 on Jurkat-76 cells expressing Vγ9Vδ2 TCR (WT), Vγ9Vδ2 TCR (TMα), and Vγ9Vδ2 TCR (AAA). Numbers in plots indicate percent of gated events. g, The characteristic peaks of cholesterol in the Vγ9Vδ2 (TMα) TCR–CD3 and Vγ9Vδ2 (AAA) TCR–CD3 complex sample. Data are representative of two independent experiments.