Isolation of cell surface proteins was achieved using Cell Surface Protein Isolation Kits (Abcam). Briefly, CD8+ Treg cells were incubated with sulfo-NHS-SS-biotin for 30 minutes and quenched for 5 minutes. Cells were then lysed and centrifuged at 10,000g for 2 minutes. Supernatant was incubated with streptavidin beads for 1 hour. Streptavidin beads were collected and washed with lysis buffer, followed by incubating with elution buffer for 30 minutes to release the proteins for subsequent analysis.

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