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. 2022 Jun 29;8(26):eabo0183. doi: 10.1126/sciadv.abo0183

Fig. 5. CpG motif analyses indicate that FoxP3 DNA methylation is not regulated by NAC1.

Fig. 5.

Naive WT and NAC1−/− CD4+CD25+ Tregs were purified and used in these analyses. (A) Diagram of mouse FoxP3 DNA methylation. Seven CpG sites of FoxP3 regulators (ADS657, ADS569, ADS1183, ADS442, ADS443, ADS779, and ADS1184) were analyzed. (B) Four CpG regions of FoxP3 regulators, including Distal Region (Regions ADS657 and ADS569), Proximal Region (ADS1183), CNS 2 Region (TSDR ADS443), and 3′ Downstream (ADS1184), were analyzed. (C) Genomic DNA of the lymphocytes from LNs and spleen of WT or NAC1−/− mice was analyzed for methylation status of CNS2 (TSDR) and FoxP3 proximal promoter, respectively. The degree of methylation at each CpG motif is depicted according to the color code. A representative is shown. (D) Percentage of methylation of each FoxP3 CpG based on two experiments. P > 0.05; unpaired t test.