Fig. 6.

mLN SCs instruct resDCs with Treg-inducing properties. Indicated LNs were transplanted to the popliteal fossa of SPF-housed mice. Eight to sixteen weeks later, single cell suspensions were generated by enzymatic digestion, and respective DC populations isolated and co-cultured with naïve CPDviolet-labeled cells isolated from Foxp3^hCD2xRag2^−/−xDO11.10 mice and 160 ng/ml Ova_323-339 peptide for five days. a Representative dotplots of Foxp3 expression over CPDviolet dilution on gated CD4⁺ T cells after co-culture with indicated DC subsets. Numbers indicate frequencies in gates. (b–c) Scatterplots show frequencies of Foxp3⁺ Tregs among CD4⁺ T cells after co-culture with migDCs (b) or resDCs (c) obtained from indicated locations. Data pooled from two to three independent experiments (n = 3–7). d–e resDCs were isolated from indicated LNs. RNA-seq^L and subsequent analysis was performed. d GO analysis of biological processes of genes persistently up-regulated (Maintained) or down-regulated (Repressed) in resDCs isolated from transplanted mLN-SPF. e Heatmap of expression of indicated genes. Numbers in brackets indicate average log₂(RPKM) expression of respective genes across all experimental groups. f Scatterplots show frequencies of Foxp3⁺ Tregs among CD4⁺ T cells after co-culture with resDCs obtained from indicated locations with or without supplementation of exogenous Noggin (100 ng/ml). Data pooled from two to three independent experiments (n = 3–7). migDC, migratory dendritic cell (Lin⁻CD11c⁺MHCII^high); resDC resident dendritic cell (Lin⁻CD11c^highMHCII⁺); Lin CD3⁺CD45R⁺Ly6G⁺F4/80^high, Endo endogenous, Tx transplanted