Extended Data Figure 10. Isotype Control Staining and BTN3A1 blocking.

(a) G115 clone Vγ9Vδ2 TCR tetramer staining MFI of WT Daudi-Cas9 cells treated with 80 μM C991 (DMSO), DMSO (vehicle), 0.5 mM AICAR (aqueous), or without treatment for 72 hours. Two-tailed unpaired Student’s t test. (b) Vγ4Vδ1 TCR (clone DP10.7) tetramer staining fluorescence (MFI) of Daudi-Cas9 KO cells treated with 80 μM C991 (DMSO), DMSO (vehicle), 0.5 mM AICAR (aqueous), or water for 72 hours. This staining with a tetramer of an irrelevant γδTCR clone defines the background for Vγ9Vδ2 TCR tetramer staining in Figure 4a. (c) qPCR data for BTN2A1, BTN3A1, and BTN3A2 transcripts in Daudi-Cas9 cells treated with C991, internally normalized to ACTB transcripts and normalized to DMSO (vehicle)-treated cells. Two-tailed unpaired Student’s t test. (d) IgG1κ isotype control staining in Daudi-Cas9 KO cells treated with 80 μM Compound 991 (DMSO), DMSO (vehicle), 0.5 mM AICAR (aqueous), or water (vehicle) treatment for 72 hours. (e) Survival of eGFP+ Daudi cells treated for 3 days with AICAR or water prior to co-culture (E:T 2:1) with primary Vγ9Vδ2 T cells in the presence of an anti-BTN3A antibody (clone 103.2). Cells were quantified using real-time quantitative live-cell imaging (Incucyte). Survival was normalized to Daudi cells cultured without T cells. (a) n=4 per condition, representative data from one of two independent experiments. (b) n=3 per condition, representative data from one of two independent experiments. (c) n=4 per condition, representative data from one of three independent experiments. (d) n=3, representative data from one of two independent experiments. (e) n=4 per condition. (a-e) Mean ± SD. p<0.0001 (****).

Source Data files provided for Figures 2, 3, 4, and Extended Data Figures 1, 4, 7, 8, 9, and 10.