Figure 5. Promoter and CpG regions are the major sites where methylation is regulated by DNMT1. (A) Intratumoral T-cells were purified by CD4-based flow cytometry from the HCC-bared mice that had received intratumoral injection with AAV-pCD4-DNMT1, or AAV-pCD4-scrambled, or AAV-pCD4-shDNMT1. (B) RT-qPCR for DNMT1 in CD4+ cells from AAV-pCD4-DNMT1-, or AAV-pCD4-scrambled-, or AAV-pCD4-shDNMT1- injected tumor. (C) % CD4+CD25+Foxp3+ Treg cells in total CD4+ cells. (D–F) Methylation status of 3 known sites on Foxp3 (promoter (D), enhancer (E) and CpG (F) region) were assessed by MS-PCR. *p<0.05. NS: non-significant. N=3.