1. Corticotropin-Releasing Hormone Stimulates Angiogenesis and Epithelial Tumor Growth in the Skin 
Jack L. Arbiser, Katia Karalis, Akila Viswanathan, Chieko Koike, Bela Anand-Apte, Evelyn Flynn, Bruce Zetter, Joseph A. Majzoub 
Journal of Investigative Dermatology 
Volume 113, Issue 5, Pages (November 1999)
DOI: /j x
Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Effect of CRH on endothelial chemotaxis in vitro. (A) Dose–response effect of CRH upon bovine capillary endothelial cell migration. A significant increase in migration is found at concentrations of CRH ≥ 10 nM (*p < 0.05 vs 0 CRH). (B) Checkerboard assay to differentiate chemotaxis from chemokinesis to CRH. The concentrations of CRH in the lower/upper Boyden chambers are denoted by the first/second numbers, respectively. The increase in migrated cell number at 20/0 nM CRH in the single lower chamber is significant (*p < 0.05 vs 0 CRH), whereas migration of endothelial cells towards CRH is abolished when equal concentrations of CRH (either 2/2 nM or 20/20 nM) are present in both chambers. (C) Effect of CRH on vascular smooth muscle cell migration. Smooth muscle cell are significantly chemotactic towards platelet-derived growth factor, 400 nM (*p < 0.05 vs 0), but not towards CRH, 2–200 nM. (D) Effect of the CRH antagonist, α-helical CRH9-41, on endothelial cell migration. The number of migrating cells in the presence of CRH (20 nM) is significantly greater (*p < 0.05) than compared with control or CRH antagonist alone (CRH Ant). Co-addition of CRH, 20 nM and CRH antagonist, 200 nM (CRH/CRH Ant) abolishes CRH-specific chemotaxis.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Growth of CRH+ and CRH– tumors in vivo. (A) Photograph of tumors in nude mice inoculated with tumor cells 4 wk previously. The mouse on the left was inoculated with CRH+ (2 × 106 cells), which overexpress the CRH and the neomycin resistance genes, whereas the mouse on the right was inoculated with CRH– (4 × 106 cells), which express the neomycin resistance gene alone. (B) Expression of tumor CRH peptide in vivo. Positively stained cells have brown cytoplasm and blue nuclei, whereas negative cells only have blue nuclei. Tumors derived from CRH+ cells express CRH peptide in their cytoplasm (left panel), whereas those arising from CRH– cells are immunonegative (right panel). (C) Growth curves of CRH+ and CRH– tumors in nude mice in vivo. At each time point, CRH+ tumor volume (closed bars) was significantly greater than CRH– tumor volume (open bars). (D) Growth curves of CRH+ and CRH– cells in vitro. Ten thousand cells of each line were plated and counted after 72 h in cell culture. CRH+ cells grew significantly more slowly than did CRH– cells in vitro.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Vascularity of CRH+ and CRH– tumors in vivo. (A) von Willebrand factor staining of CRH+ and CRH– tumors, excised 2 wk after implantation. Positively stained blood vessels appear red. (B) Microvessel count per high power (200×) field. Tumors arising from CRH+ cells had a significantly greater number of von Willebrand-positive blood vessels than did tumors arising from CRH– cells.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions