Figure 2

Genetic targeting of pan-endothelial heparan sulfate biosynthesis impairs metastasis of carcinoma to regional lymph nodes. LLC tumor cells were injected subcutaneously into the left caudal/medial inguinal region of Ndst1^f/fTekCre+ mutant mice, which bear a pan-endothelial mutation in the major HS sulfating enzyme Ndst1, and their wildtype Cre- littermates as controls. After 14 days, the left subiliac lymph node (draining the primary tumor) from each mouse was isolated. A. Metastaic tumor cells in the lymph nodes (LN) were detected using anti-pan-keratin antibody (blue stain) on LN tissue sections imaged under 100× magnification, and quantified (NIH Image-J) to determine net pixel intensity for each LN. Values were normalized to the mean pixel value/LN among control (Ndst1^f/fTekCre-) littermates. Graph is shown below; *P = 0.016, for difference in mean values (red bars) for mutant (N = 8) vs control (N = 6) mice. B. Representative LN tissue sections from tumor-bearing mutant (bottom row of photomicrographs) vs wildtype (Cre^-) controls (middle row) were co-stained with CCL21 (blue) and LYVE1 (brown) (left two panels) or pan-keratin (blue, right panel). The top row of photomicrographs shows a representative LN section from a non-challenged (tumor free) control mouse. Scale bars, 100 μm for 100× and 50 μm for 400× magnification photomicrographs.