Soluble antigen dispersal and DC-mediated uptake in LNs. (A) Mice were injected with EαGFP in the footpad, and draining popliteal LNs were isolated at the indicated time points for visualization of antigen dispersal, as well as uptake by LN-resident DCs. (I) Visualization of the overall EαGFP dispersal across LNs. (II) Identification of stromal elements within LNs, with Lyve-1 (white) signal demarcating the LS, Meca79 (red) staining highlighting the HEVs, and collagen-IV (ColI.IV, blue) staining predominantly denoting the LN conduits. Accompanying gp38 staining of conduits is shown in Fig. S1. (III) Quantitative heat-map analysis of EαGFP signal within conduits, as generated by creating surface objects around LN conduits and quantifying mean EαGFP fluorescence. (IV) Localization of CD11c^HIGHMHC-II^INTCD169⁻ LN-resident SIRPa⁻ cDC1 (cyan) and SIRPa⁺ cDC2 (red), with respect to B cells (blue) and CD169⁺ macrophages (yellow). (V) Quantitative heat-map analysis of EαGFP signal within DCs (EαGFP signal gated within CD11c⁺Coll.IV⁻Lyve1⁻Meca79⁻gp38⁻ voxels). Bars, 200 µm. (B) Conduits (top) and DCs (bottom) from A were quantified for total EαGFP content with respect to distance from the nearest LS. Data represent at least two independent experiments. Numbers in plots represent frequencies.