A total of 125 pairs of CRC samples and adjacent normal tissues were obtained from CRC patients at the First Affiliated Hospital of Soochow University (Suzhou, China). All experiments involving patient specimens were approved by the Institutional Review Board of Soochow University. Informed consent was obtained from each patient. Detailed clinicopathological information is provided in Supplementary Table S1 . An IHC assay was conducted as previously described17 (link). Briefly, CRC samples or xenograft tumor samples were deparaffinized and rehydrated. After antigen retrieval with 10 mM sodium citrate buffer (pH 6.0), the sections were incubated with goat anti-human 4IgB7-H3 antibody (R&D Systems, MN, USA, #AF1027, 1:100), mouse anti-human CD31 antibody (Abcam, Cambridge, MA, USA, #ab32457, 1:1500), or rabbit anti-human VEGFA antibody (Proteintech, Wuhan, China, #19003–1-AP, 1:500). All sections were then reviewed blindly by two experienced pathologists (Dr. Cao and Dr. Zhan). The scoring criteria for IHC staining were based on the intensity of immunostaining and the percentage of immunoreactive cells as described previously17 (link). We calculated the score of the MVD on the basis of previously described criteria22 (link).
Free full text: Click here
