Fig. 4

JunB regulates expression of eTreg-related molecules. a Flow cytometry analysis of CD4⁺Foxp3⁺ Treg cells in various tissues isolated from Cd4^CreJunb^fl/fl and Junb^fl/fl mice. Graphs show percentages and numbers of CD4⁺Foxp3⁺ Treg cells. Error bars indicate s.d. (n = 4). *P < 0.05 (unpaired two-tailed Student’s t test). b, c Flow cytometry analysis of CD44 and CD62L in CD4⁺Foxp3⁺ Treg cells isolated from spleens of Cd4^CreJunb^fl/fl and Junb^fl/fl mice (8–12-week-old). Graphs show percentages of CD62^hiCD44^lo cTreg and CD62^lo eTreg cells b, and MFIs of CD44 c. Error bars indicate s.d. (n = 4). *P < 0.05 (unpaired two-tailed Student’s t test). d, e Flow cytometry analysis of CTLA4, CD25, and GITR d, and ICOS, TIGIT, KLRG1, and ST2 e in CD62^hiCD44^lo cTreg cells and CD62^lo eTreg cells among CD4⁺Foxp3⁺ Treg cells isolated from spleens of Cd4^CreJunb^fl/fl and Junb^fl/fl mice (8–12-week-old). Representative flow cytometry profiles show CD62^hiCD44^lo cTreg cells and CD62^lo eTreg cells d, and CD62^lo eTreg cells e. Graphs show MFIs of CTLA4, CD25, and GITR d, and percentages of cells expressing indicated molecules e. Error bars indicate s.d. (n = 4). *P < 0.05; N.S., not significant (unpaired two-tailed Student’s t test). f) Flow cytometry analysis of ICOS in Nrp1⁺ and Nrp1⁻ Treg cells isolated from spleens of Cd4^CreJunb^fl/fl and Junb^fl/fl mice (8–12-week-old). The graph shows percentages of ICOS-expressing cells among Nrp1⁺ and Nrp1⁻ Treg cells. Error bars indicate s.d. (n = 4). *P < 0.05 (unpaired two-tailed Student’s t test). g Flow cytometry analysis of ICOS, TIGIT, and KLRG1 in CD4⁺Foxp3⁺ Treg cells isolated from spleens of 1-week-old Cd4^CreJunb^fl/fl and Junb^fl/fl mice. Graphs show percentages of cells expressing indicated molecules among CD4⁺Foxp3⁺ Treg cells. Error bars indicate s.d. (n = 3 for Junb^fl/fl, n = 7 for Cd4^CreJunb^fl/fl). *P < 0.05 (unpaired two-tailed Student’s t test). Data represent two independent experiments