We cocultured Tregs in transwells with brain slices collected 5d after tMCAO in the lower compartment for 24h to activate Tregs. Tregs in the transwells were transferred to new wells with primary cultured microglia, and cocultured for 2d. Microglia were then collected for bulk RNAseq analyses. The results revealed roles of activated Tregs in polarizing microglia toward an anti-inflammatory and reparative phenotype. Overall design: Cocultured Tregs in transwells with brain slices collected 5d after tMCAO in the lower compartment for 24h. And then Tregs in the transwells were transferred to new wells with primary cultured microglia, and cocultured for another 2d. Microglias stimulated with or without Treg were send to bulk RNAseq.