Blockade of VEGF-A profoundly attenuates the LPS- and LTA-induced lymphatic vessel densities in the inflamed ear skin and DLNs, and lymphatic flow and mobilization of inflammatory cells from the inflamed skin to DLNs. VEGF-Trap (25 mg/kg), dimeric-Fc (Fc, 25 mg/kg), or none was treated at 1 day before and after the intradermal injection of PBS (P), LPS, or LTA. (A) At day 3 after the injection, the inflamed ears and DLNs were sectioned for histologic analysis. Tissue sections were coimmunostained for LYVE-1 and CD31. Scale bars represent 50 μm. (B,C) Lymphatic (LV) and blood vessel (BV) densities in the inflamed skin (B) and DLNs (C) were quantified and presented as percentages. (D,E) Three days later, 3 μL FITC-conjugated dextran was intradermally injected into the inflamed skin. Thirty minutes later, fluorescence intensities in DLNs were determined with the IVIS (top panels) and fluorescence stereomicroscope (bottom panels), and quantified and presented as relative radiance (photons/sec per cm²/steradian). (F) At day 3 after intradermal injection of LPS or LTA into the ear, the GFP⁺ inflammatory cells (∼ 10⁶ cells) were injected intradermally into the inflamed skin. Twelve hours later, the DLNs were sectioned and immunostained for LYVE-1. Scale bars represent 50 μm. (G) The GFP⁺ inflammatory cells in the section of DLNs are quantified and presented as AU. All bars represent mean ± SD from 4 to 5 mice. *P < .05 versus P; ^#P < .05 versus LPS + Fc or LTA + Fc.