Fig 1: Loss of spatial vicinity of DCs and HEVs prohibits LTßR signaling(A) FCM analysis of surface molecules in HECs from LNs of Ltbr+/+ and Cdh5CreERT2-Ltbrfl/fl mice (Ltbrfl/fl). Gating strategy given in Figure S5B.(B) Lymphoma-bearing mice treated with either isotype antibody or stimulatory anti-LTßR antibody. Quantitation of PNAd+ cells among BECs and ICAM-1 expression intensity in these cells as percentage of controls. n = 8 mice per group.(C) Quantification of LN infiltrating adoptively transferred lymphocytes by FCM.(D) Analysis of migratory DCs (mDCs) and classic DCs (cDCs). Left: proportion of DCs among all CD45+ cells. Right: total DC numbers per LN. Control n = 10, tumor n = 6 mice.(E) Top: confocal microscopy images of DCs in LNs of Cd11cGFP mice with and without tumor. Bottom: segmented images to highlight cell-cell contacts.(F) Representative images of DC distribution in LN sections of Cd11cGFP mice. Boxes indicate magnified and segmented areas.(G) Density of DCs (Cd11cGFP) among the FRC scaffold (PDPN).(H) Quantification of the proportion of the DC network area (determined by Cd11cGFP/MHC-II expression) relative to the area of the whole LN section in percent. n = 5–6 mice per group.Scale bars, 10 µm (E and G) and 100 µm (F). Mean and SEM are indicated. All data points represent individual animals. Statistics were calculated with Mann-Whitney U-test. *p < 0.05; **p < 0.01; ***p < 0.005; n.s., not significant.