Extended Data 4. Gating strategy to measure γδ2 T cell degranulation and parasite reinvasion.

a, To measure degranulation, γδ2 T cells were co-cultured with RBCs in the presence of anti-CD107a for 4 hr. Cells were then stained with viability Live/Dead dye and anti-CD3. Single live cells were gated on SSC-A vs FSC-A, excluding dead cells. CD107a⁺ staining was analyzed on gated CD3⁺ γδ2 T cells. b, To determine the effect of γδ2 T cells on parasite reinvasion, synchronized iRBCs infected 12, 30 or 42 hr earlier were cultured for 42, 24 and 12 hr, respectively, with or without γδ2 T cells at different E:T ratios. Parasite reinvasion was measured by flow cytometry using SYBR green staining to detect parasite DNA and anti-CD235a for RBC gating and anti-CD3 to exclude γδ2 T cells. The DNA content of iRBCs at different stages enabled gating on each stage of parasite infection to quantify the proportion of iRBCs at each stage. Reinvasion of fresh RBC increased the proportion of rings. The reinvasion % was calculated as the percentage of newly invaded RBCs at ring stage in comparation with the Plasmodium culture without γδ2 T cells or any treatment (100% reinvasion).