Fig 2: Tumor necrosis factor receptor signaling regulates Podxl and Cd300lg expression in PNAd+ TEC. C57BL/6 WT or TNFR1/2-/- mice were I.P. injected with B16-OVA cells and tumors were harvested 14 days after implantation. LN were harvested from non-tumor-bearing mice. (A, B) Endothelial cells were sorted from CD45+ depleted LN and I.P. tumor suspensions, and the expression levels of indicated RNA transcripts were determined by qPCR, as described in Methods. Data from one experiment presented as 2-?CT relative to Hprt, n=5 LN or I.P. tumors per group. (A-B) Results are mean ± SD analyzed by unpaired Welch’s t-test. *p<0.05, **p<0.01, all other paired bar comparisons are not significant (p>0.05) and have been omitted for clarity.

Fig 3: Scaffolding proteins are expressed at significantly lower levels in PNAd+ TEC than in LN PNAd+ BEC. C57BL/6 mice were I.P. injected with B16-OVA cells and tumors were harvested 14 days after implantation. LN were harvested from non-tumor-bearing mice. Endothelial cells were sorted from CD45+ depleted LN and I.P. tumor suspensions, and the expression levels of indicated RNA transcripts were determined by qPCR, as described in Methods. Data from one experiment presented as 2-?CT relative to Hprt, n=5 LN or I.P. tumors per group. Results are mean ± SD analyzed by unpaired Welch’s t-test. **p<0.01, all other paired bar comparisons are not significant (p>0.05) and have been omitted for clarity.

Fig 4: Balb-ADAM10B-/- LN exhibit WT architecture unlike C57-ADAM10B-/- nodes.Naïve LN sections from C57Bl/6 and Balb/c WT and ADAM10B-/-; (A) B cell (blue, B220), T cell (red, Thy1.2), and HEV (green, pNAD); (B) FDC reticula (red,CR1/2), collagen (green), and cortico-medullary junction (dotted line in inset box); (C) TNF staining (green), B cell (blue, B220) follicle. (D) Average TNF staining intensity representing 12 follicle sections per group. Scale bar, 50µm. **p<0.005.

Fig 5: Glycosyl transferases are expressed at significantly lower levels in PNAd+ TEC than in LN PNAd+ BEC. C57BL/6 mice were I.P. injected with B16-OVA cells and tumors were harvested 14 days after implantation. LN were harvested from non-tumor-bearing mice. Endothelial cells were sorted from CD45+ depleted LN and I.P. tumor suspensions, and the expression levels of indicated RNA transcripts were determined by quantitative PCR (qPCR), as described in Methods. Data from one experiment presented as 2-?CT relative to Hprt, n=5 LN or I.P. tumors per group. Results are mean ± SD analyzed by unpaired Welch’s t-test. **p<0.01.