Figure 4.

An expansion/de novo conversion of Tregs occurs in peripheral organs at days 5 and 10 of pregnancy. Rag-1^-/- female mice were transferred with CD4⁺Foxp3^GFP- cells from Foxp3^GFP mice and CD4⁺CD25⁺ cells from normal C57/BL6 WT mice were added to avoid autoimmunity. After allowing reconstitution for a period of 4 weeks, the animals were paired with BALB/c males and possible conversion of CD4⁺Foxp3^- into CD4^-Foxp3⁺ cells was traced by de novo expression of CD4⁺Foxp3^GFP Tregs in samples of reconstituted mice at days 5 (d5) and 10 (d10) of pregnancy (A) in the draining lymph nodes (B and C), mesenteric lymph nodes (D), blood (E), inguinal lymph nodes (F) and spleen (G) (n=3-5). Non-pregnant reconstituted animals were used as a control. C shows representative dot plots of the analyzed frequency of CD4⁺Foxp3^GFP+ cells in draining lymph nodes from one animal representative of each group. Data are expressed as single dots with medians and were analyzed by using the one way ANOVA, Kruskal-Wallis test, followed by a Dunn’s multiple comparison test (*: P <0.05).