Figure 4.

Potential negative selection and Treg development in sTOs

(A) Representative IF image of AIRE in week 4 sTOs (n = 2).

(B) Representative IF image of PD-1 and CD3 in week 4 sTOs (n = 2).

(C and D) FC plots (C) and quantification (D) of PD-1 expression in CD45⁺/CD3⁺ cells in 1$°$ thymi and week 4 sTOs (thymi, n = 2; sTOs, n = 3 ind. exp., 1 hPSC line).

(E and F) FC plots (E) and quantification (F) of FOXP3 expression in CD25⁺/CD4⁺ cells in 1$°$ thymi and sTOs (thymi, n = 3; sTOs, n = 6 ind. exp., 2 hPSC lines).

(G and H) qPCR analysis of FOXP3 (G) and TRAs (H) in 1$°$ thymus, TEPs, and week 4 CD45⁻ sTO cells (G: thymi, n = 3; TEPs, n = 6–7 ind. exp., 4 hPSC lines; sTOs, n = 5–6 ind. exp., 4 hPSC lines; H: TEPs, n = 5 ind. exp., 1 hPSC line; sTOs, n = 3 ind. exp., 1 hPSC line).

Plots show percentage of the mean; data are shown as mean ± SEM; p values were determined by t test.