Anti pecam1 cd31

Confluent cultures of WT and MT-HGF primary keratinocytes were used for grafting as described previously (30 ). Six million keratinocytes (WT or MT-HGF) were respectively mixed with 6 million WT or MT-HGF mouse primary dermal fibroblasts (cultured for 1 week) and grafted onto syngeneic recipient back (WT or MT-HGF) on a prepared skin graft site located in the interscapular region. Three to four weeks post-grafting when tumors had developed on the MT-HGF grafts, daily gavage treatment with gefitinib (100mg/kg) or vehicle control (10% DMSO in water) was conducted for 2 weeks. This dosage and regimen was chosen based on efficacy and lack of side effect as reported previously (42 (link)). Tumor dimensions were measured weekly using calipers, and approximate tumor volumes were determined by multiplying tumor height × length × width. Tumors were collected in 10% formalin and processed for H&E staining (Histoserv Inc.), Ki67 and or anti-PECAM-1/CD31 (Santa Cruz, sc-1506) immunostaining was performed by the Pathology/Histotechnology Laboratory (PHL, NCI-Frederick). The percentage of Ki67-positive cells was quantified by counting Ki67-labeled basal cells from at least three randomly chosen areas. The microvascular density was determined by counting the number of CD31 stained cells in at least five fields (x400 magnification, Nikon Eclipse E400).