Treg-specific hypomethylation at the TIGIT locus. (A) DMPs and RDM in TIGIT locus. (B) Location of the PCR fragments neighboring DMPs in TIGIT (biF1-5) and FOXP3 loci and clonal bisulfite sequencing results. Ref. seq, reference sequence; Bis. seq, clonal bisulfite sequencing fragments. (Arrows) DMPs identified from 450k array. Data are represented as the mean of 5 donors. (C) Methylation-sensitive Taqman-qPCR showing methylation at TIGIT (RDM) and FOXP3 (TSDR) loci in Act-Naive and iTreg and demethylation in Act-rTreg. Data are represented as mean + standard deviation of 3 donors; paired t test (2-tailed). (D) Expression of TIGIT and CD25 in Act-Naive, iTreg, and Act-rTreg. Data are representative of 10 healthy individuals all showing similar results.