1. Vascular HR ligand expression depends on tumor anatomical location.
Vascular HR ligand expression depends on tumor anatomical location. A–I, B16-OVA tumors were harvested 14 days after SC injection or 11 days after IP injection. Ears (H) and intestine (I) were harvested from non–tumor-bearing mice. Samples were digested, enriched for CD31+ ECs, and analyzed by flow cytometry, or frozen, and processed for immunofluorescence microscopy. (A) Representative and summary data of CD31+CD45neg ECs stained for E-selectin (n = 3 tumors per group, 3 independent experiments), (C) P-selectin (n = 3 tumors per group, 2 independent experiments), and (F) intracellular CXCL9 (n = 7 tumors per group, 2 independent experiments). Representative and summary data (5–10 random fields from 1 section each of 3 tumors) of tumor sections costained for (B) CD31 and MadCAM-1 (3 independent experiments), (D) ICAM-1 (2 independent experiments), or (E) HA (3 independent experiments). G, VCAM-1 expression was determined either with or without tyramide amplification (n = 3 tumors per group, 4 independent experiments). Percent positive pixels determined using amplified signal. H, E-selectin and CXCL9 expression on CD31+CD45neg ECs from skin and SC tumor was determined by flow cytometry (n = 3 samples per group, 2 independent experiments). VCAM-1 expression on CD31+ pixels from skin and SC. tumor was determined by immunofluorescence (5–10 random fields from 1 section each of 3 tumors, 2 independent experiments). I, MadCAM-1 and VCAM-1 expression on CD31+ cells from colon and IP tumor was determined by immunofluorescence (5 random fields from one section each of 2 colons and 5–10 random fields from 1 section each of 3 tumors, 2 independent experiments).
Amber N. Woods et al. Cancer Immunol Res 2017;5:
©2017 by American Association for Cancer Research