Fig 2: EPZ004777 and Thiostrepton improved BMDCs maturation in vivo. (A and B) Expressions of CD86, MHC-II, CCR7, and PD-L1 on gated CD11c+ cells in BMDCs were assessed by FACS. (C and D) The expressions of CD3, CD8, and CD4 on gated CD3+ T cells, and Foxp3 and CD25 on gated CD4+ T cells from spleen were assessed by FACS. The amounts of CD3+/CD4+ cells set at 10 000. A/C is from C57BL/6J, B/D is from BABL/c mice, each group contained at least eight mice. Data were shown as means ± SD from at least three independent experiments. # P < 0.05, ## P < 0.01 compared with the model.

Fig 3: Insufficient iron contributes to Treg cell expansion. 3-weeks old female C57 mice were fed with ND or LID for 5 weeks and then were sacrificed for analysis. (A, B) qPCR of iron-related gene Tfrc and Fth1 (A) and Treg cell-related gene Il2r and Foxp3 (B) in splenic CD4+T cells of ND- and LID-treated mice. (C) Representative flow cytometry of Treg cells gated on CD4+T cells in dLNs and spleen of ND- and LID-treated mice. (D, E) quantification of the percentage and numbers of Treg cells in (C). **P < 0.01, *P < 0.05 (unpaired two-tailed Student’s t-test for A, B, D, E). Data are shown as mean ± S.E.M. Two independent experiments were performed, n = 4.

Fig 4: Changes of stimulated CD4 T subsets and signals when stimulations co-incubated with CD8 T(A) UMAP projection of CD4 T cells including rest T, stim T, and stimCo T, colored by T cell subset.(B) Differences of cell distributions of rest T, stim T, and stimCo T, with HSPhi T and IFNhi T marked by circles.(C) Fraction of T subsets in rest T, stim T, and stimCo T. The lines between two bars link the T cell subset counterparts in different samples.(D) Cell composition of T cell subsets based on cell sources.(E) DEGs between stim T and stimCo T. Also see Data D4.(F) DEGs of IFNhi between stim T and stimCo T.(G) Expression level of stim T-specific genes and stimCo T-specific genes in activated T cell subsets.(H) Expression levels of CXCR4 and IL2RA in T cell subsets of stim T and stimCo T. (I) FACS analyses of CD25 in stim T and stimCo T.(J) FACS analyses of CD25 in stimulated T derived from CXCR4low T and stimulated T derived from CXCR4low T in the presence of CD8 T. Also see Figures S4 and S5.