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. 2018 Oct 1;215(10):2509–2519. doi: 10.1084/jem.20172082

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© 2018 Tanaka et al.

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Figure 5. — Sox12 binds to and activates the promoter of Foxp3. (A) Naive CD4⁺ T cells were infected with retrovirus of pMX-IN-HA-Sox12, and cells were restimulated with TCR for additional 12 h. ChIP-qPCR assay for the promoter and CNSs of Foxp3 gene locus was performed with anti-HA antibody or control rabbit IgG. Data are compiled from three independent experiments. (B) Luciferase assay with indicated Foxp3 promoter Luci constructs in EL4 cells. Data are compiled from three independent experiments. (C) DNA precipitation assays of HA-Sox12 with biotinylated double-stranded DNA probes containing a portion of Foxp3 promoter or its mutant at Sox12-binding site (ΔACCAAAG) were performed. FXO⁺ probe was used as a positive control. Shown are representative of three independent experiments. (D) Luciferase assays with pGL4.23-Foxp3 CNS1 enhancer-Luci construct or pGL4.23 (as a control) were performed in EL4 cells. Data are compiled from four independent experiments. (A and B) *, P < 0.05; ***, P < 0.001 by unpaired t test.