Gating strategy and activation of PBMCs. (A) Representative flow cytometry dot plots of proliferating CD3⁺MR1−5-OP-RU tet⁺CTV^lo MAIT cells related to Fig. 4 A. (B) Shown in B is the gating strategy used in PBMC experiments. (C) Human PBMCs were titrated with indicated ligands for 16 h, and then, CD69 MFI was measured on the surface of gated MAIT cells. (D) PBMCs were titrated with lumichrome in the presence of zoledronate, and then, % CD69⁺ Vδ2⁺ cell population was measured. Data in C and D represent the average of three independent experiments performed in duplicate on PBMCs from three different donors. The error bars represent the standard error of the mean (SEM). One-way ANOVA statistical analysis was performed for all samples using Dunnett’s multiple comparisons (ns: not significant, ****P < 0.0001).