In vitro suppression assay and Treg culture
Responder cells were labelled with 6 µM Cell trace violet (Thermo
Fisher), then cultured with or without unlabeled Treg cells at a 1:1
ratio for 72 h in the presence of Dynabeads Mouse T-activator CD3/CD28
(1 bead/cells) in round-bottomed 96-well dishes. The proliferation
inhibition rate on the responder was calculated as: (1-(Cell trace
violet percentage Treg plus responder cells co-culture/responder cells
alone)) x 100%.