Fig 2: Developing T cells at the ß-selection checkpoint establish an immunological synapse(Ai and Aii) Representative images acquired using Vectra imaging system show a 6 color multiplex panel (Ai) used to show that developing T cells (CD25+ CD44- CD4- CD8-) establish an immunological synapse (Aii, bottom right, red arrow) as indicated by the co-clustering of the pTa subunit with its downstream effector pLCK394 the immunological synapse. Scale Bar; Ai) 100 µm; Aii) 10 µm.

Fig 3: Confirming specificity of CD44 and CD4+CD8 before co-staining using OPAL 650(A) Representative images of dual multiplex staining of CD25, CD44 and DAPI acquired using Vectra imaging system showing CD25+ CD44- cells (green arrow head, zoomed image), CD25- CD44+ cells (red arrow head, zoomed image) and CD25+ CD44+ cells (yellow arrow head, zoomed image).(B) Representative images of dual multiplex staining of CD25, CD44 and DAPI acquired using Vectra imaging system showing CD25+ CD44- cells (green arrow head, zoomed image). Scale Bar; A&B) 100 µm; A&B) zoomed images 10 µm.

Fig 4: Colon and MLN immune cell changes in splenectomised mice. There was no difference between the groups when analysing the colons for (A) CD4+, (B) CD8+, (D) F4/80+ and (E) B220+ cells. TUS induced an increase in (C) colonic CD25+ T cells while decreasing the percentage of (H) CD25+ T cells in MLN. Furthermore, (F) CD4+ T cell levels were decreased in the MLN, while no difference was seen regarding (G) CD8+, (I) F4/80+ and (J) B220+ cells. *p < .05 compared to 2% DSS. Student's t-test. N = 4/group. Results are presented as mean ± SD.