A. Single cell suspensions prepared from different anatomical compartments of the small bowel (SB) including the IEC fraction, lamina propria (LP), serosa with the intestinal muscularis and PP were analyzed by seven color flow cytometry. The results were compared to the whole SB depleted of PP (SB without (w/o) PP). To define CD11chi and CD11clo DCs, gates were set on viable DAPI−CD45+MHCIIhi cells. CD11chi DCs were further divided into CD103+CD11b−, CD103+CD11b+ and CD103−CD11b+ DCs as indicated in the dot plots. Dot plots in the left column show the % of CD11chi and CD11clo DCs among total DAPI−CD45+MHCIIhi cells. Dot plots in the right column show the % of CD103+CD11b−, CD103+CD11b+ and CD103−CD11b+ DCs among total MHCIIhiCD11chi or MHCIIhiCD11clo (serosa and muscularis) DCs. B. Images show the distribution of CD103+CD11c-YFP+ cells in the villi of distal SB from CD11c-EYFP transgenic mice. Magnification 400x. Scale bar 10 nm. C. Dot plots show the % of CD103+CD11b−, CD103+CD11b+ and CD103−-CD11b+ DCs among CD45+MHCIIhiCD11chi cells in the SB depleted of PP in WT mice and in the total SB of Id2−/− mice. D. Overlay histograms show the differential MHCII, CD8α, CX3CR1, M-CSFR and F4/80 expression among each intestinal DC subset. E. Images show purified CD103+CD11b+ and CD103−CD11b+ SB LP DCs spun onto glass slides and stained with Giemsa. Magnification 600x. Scale bar 10 nm. F. CFSE labeled OTII cells were cultured with purified CD103+CD11b+ or CD103−CD11b+ SB LP DCs, splenic DCs or splenic B cells pulsed with OVA-peptide. Numbers are the % of OTII cells that have not proliferated in each group (representative data of two independent experiments done in triplicates).