Integrins are required for the maintenance of cDC2s in blood-exposed regions of spleen. (A) Frequencies of in vivo anti-CD45-PE–labeled CD8⁺ cDC1s in Cd11c-Cre Talin^fl/fl and control mice. Each symbol represents one mouse, and lines denote means. One of two independent experiments with similar results is shown. (B) Total splenocytes isolated from Cd11c-YFP reporter mice (CD45.2⁺) were transferred into CD45.1⁺ mice. Representative profiles of transferred DCs in blood after 30 min and 3 h. One of two independent experiments with similar results is shown, in which each group had at least three mice. (C–E) Frequencies of DCIR2⁺ cDC2s in spleen (C) and of in vivo anti-CD45-PE labeling of cDC2s (D) or representative distribution patterns of DCIR2⁺ cDC2s (red), CD169⁺ MMMs (green), and SIGNR1⁺ MZMs (white) relative to IgD⁺ B cells (blue) in spleens 15 h after anti-α4/αL or saline treatment (E). Each symbol represents one mouse, and lines denote means. Data are pooled from two independent experiments. Scale bar, 200 µm. (F and G) Frequencies of in vivo anti-CD45-PE labeling of cDC2s (F) or representative distribution patterns of DCIR2⁺ cDC2s (red) relative to IgD⁺ B cells (blue) in spleens 4 h after FTY720 and 15 h after anti-α4/αL or saline treatment (G). Scale bar, 200 µm. (H and I) Frequencies of CXCR5^hi PD-1^hi T cells among transferred OT-II T cells (H) or FAS^hi GL7^hi GC B cells in B220⁺ B cells (I) in 4-h FTY720- or saline-pretreated mice, 6 d after SRBC-OVA immunization. (J) Representative distribution patterns of DCIR2⁺ cDC2s (red) relative to IgD⁺ B cells (blue) in spleens 5 h after soluble OVA plus LPS immunization. Scale bar, 200 µm. Each symbol represents one mouse, and lines denote means. One of two independent experiments with similar results is shown. Sections are representative of multiple spleen cross sections from six mice of each type. **, P < 0.01; ***, P < 0.001.