Fig. 1
From: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
Identification of tumor endothelial cell-specific phage clones and hornerin as a non-VEGF but TCM-induced protein. a The amino-acid sequences of 30 selected phage clones that displayed selective binding to pancreatic tumor endothelium. b Orthotopic pancreatic tumor section showing co-localization of FITC-labeled “PRH” motif phage clones (green) with PCAM1-positive vessels (red). Relative affinity and fold increase over normal (adjacent, unaffected pancreas vessels) of the “PRH” pool is presented below the image. c Venn diagram generated after phage-based ELISA and two-way ANOVA analysis that illustrates specificity of phage clone binding to VEGFCM- and/or TCM-treated HUVECs. d Quantification of the specificity (fold over vehicle control (VC)) for each clone in the Venn diagram. N = 6 for each clone. e Immunoblot detection of hornerin in lysates generated from M13KE (control; lane 1) and PTEM 9 (lane 2) phage pulldowns. f Representative immunoblot of lysate preparations from VC (lane 1), VEGFCM (lane 2), and TCM (lane 3)-treated HUVECs display the levels of hornerin protein. HSP90 is shown as a loading control. Molecular weight is indicated in kilodaltons (kDa). g Relative densitometry measured as hornerin fold expression over HSP90. N = 3 for each condition. h Immunofluorescence detection of hornerin (column 1) on non-permeabilized HUVECs following treatment with VC (top row), VEGFCM (middle row), or TCM (bottom row). Wheat germ agglutinin (column 2) was included in the stain to highlight the plasma membrane. The merged image (column 3) displays hornerin and WGA co-localization. Scale bar = 50 μm. Graphs represent the mean ± SEM. *P ≤ .05 by unpaired two-tailed t-test, treatment compared to vehicle control
