Fig. 7. TolDC treatment recruits Tregs to the lung and the spleen. The lung and spleen cell preparations obtained from mice of each experimental group were stained with anti-CD4-APC and anti-CD25-FITC, and after permeabilization the cells were stained with anti-Foxp3-PE. (A) Gating on CD4⁺ T cells, and representative dot plots of lung Tregs (CD4⁺CD25^highFoxp3⁺) and isotype controls. (B) Quantification of lung Tregs (CD4⁺CD25^highFoxp3⁺) relative to total CD4⁺ T cells. (C) CD4⁺CD25^highFoxp3⁺ splenocytes. (D) Confocal microscopy images showing CD3 (green), Foxp3 (red), and nuclei stained with DAPI (blue) in the lung tissue of TolDC-treated mice. Values are expressed as means±SEM of 5–7 mice per group. TolDC, tolerogenic dendritic cell; Treg, regulatory T cell; FITC, fluorescein isothiocyanate; DAPI, 4′,6-Diamidino-2-phenylindole; SEM, standard error of the mean. ^*P<0.05; ^†P<0.01; ^‡P<0.001 (Bonferroni's multiple comparison test).