Effects of CD80/CD86 blockade and subsequent antigen feeding on antigen-presensitized mice. (A and B) Changes in ear thickness of the right ear (A) and body weight (B) of the mice treated as shown in Fig. 7 A. Arrowheads indicate OVA challenges. (C) Representative histologies of the left ear of the mouse groups at day 33 as shown in Fig. 7 C. Bars in histology indicate 100 µm. (D–F) Experimental protocol to assess the effects of EWP doses on oral tolerance induction by CTLA4-Ig treatment and EWP feeding (D). DORe mice were OVA-sensitized at the right ear four times, i.p. injected with CTLA4-Ig on day 11, fed with 1, 10, or 30% EWP chow for 4 days, OVA-challenged at the left ear four times from day 22, and assessed for body weight (E) and ear swelling (F) (n = 4). (G) Demethylation status and frequencies of demethylation at the Foxp3-CNS2 locus of DORe-pTreg cells isolated from the mice treated with saline or 10 µg CTLA4-Ig followed by EWP feeding and assessed at day 15 in Fig. 7 A. (H and I) Frequencies of pTreg cells in mLNs, dLNs, and left ears (H) and of IFN-γ⁺, IL-4⁺, and IL-17A⁺ cells in dLNs at day 33 in DORF mice treated as described in Fig. 7 J (I) (n = 6–7). Vertical bars indicate the mean ± SEM. Statistical significance was assessed by an unpaired t test or Welch’s t test (H and I). P values were shown in the figures.