Fig 1: Desminlow/aSMAhigh phenotype is correlated with vessel leakiness and hypoxia. (A) Representative images of FFPE sections of KPC and PANC-1 tumors, and healthy pancreas immunolabeled for Desmin and CD31. Sections are counterstained with DAPI (blue) to visualize nuclei. (B) Quantification of percentage of CD31+ vessels associated with Desmin+ pericytes. (WT n = 4; KPC and PANC-1 ortho-tumors n = 3). One-way ANOVA with Dunnett’s multiple comparisons was used to determine statistical significance. (C) Correlation of FITC-dextran leakage and Desmin+ pericyte-covered vessels (n = 6). (D) Correlation of hypoxic area and Desmin+ pericyte-covered vessels (n = 6). (E) Correlation of Desmin+ pericyte and aSMA+ pericyte-covered vessels (n = 10). Pearson’s r correlation coefficient and significance levels are presented. (F) The ratio of Desmin+ pericyte and aSMA+ pericyte per each tissue section was compared between the healthy pancreas and PDAC tumors for both murine models and humans. (G) Plasma Ang2 level was compared between KPC tumor-bearing mice and healthy mice using the Ang2 ELISA kit. Unpaired two-tailed t-test was used to determine statistical significance. Scale bar: 100 µm; apply to all images. Unless otherwise stated, the data are represented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.
Fig 2: TDB treatment leads to EC activation and upregulation of adhesion molecules in vivo A, BRepresentative images of liver (A) or tumor (B) sections from the Founder5 tumor-bearing mice stained with ICAM-1 (red), VCAM-1 (white), and Endomucin (green) 24 h post treatment. Right panels are enlarged views of the boxed areas showing only ICAM-1 and VCAM-1. Scale bar = 100 µm. Below: quantification of total ICAM-1+ and VCAM-1+ areas as well as vascular (vasc.) ICAM-1+ (ICAM-1+ Endomucin+) and vascular VCAM-1+ (VCAM-1+ Endomucin+) areas normalized to tissue areas. n = 9 mice/treatment.C, DPrincipal component analysis (PCA) plots for bulk RNAseq analysis of the liver (n = 3, C) or tumors (n = 3, D) from the Founder5 model 24 h post treatment. Each dot represents a sample and is colored by treatment (left plot). Boxplots (right) show the gene set enrichment score for common adhesion markers in the liver (C) and in the tumor (D) samples. Boxes represent interquartile ranges (IQR), and medians are horizontal lines in the boxes. Whiskers above and below the boxes are the lowest and highest values within 1.5 times IQR.E, FDensities of total blood vessels (left panel) and mature blood vessels (right panel) in liver (E) and tumor (F) sections from the Founder5 tumor-bearing mice normalized to tissue areas. n = 9 mice/treatment.G, HLeft panels: representative images of liver (G) or tumor (H) sections from the Founder5 model stained with ICAM-1 (white), pericyte markers (pink), and Endomucin (green) (top panel) or stained with VCAM-1 (white), pericyte markers (pink), and Endomucin (green) (bottom panel) 24 h post treatment. Scale bar = 100 µm. Right panels: linear regression analysis comparing adhesion molecule (ICAM-1 and VCAM-1) expression (x-axis) and vessel maturity (y-axis). The coefficients of determination (R) are indicated. n = 9 mice/treatment.I–KQuantification of circulating (I, n = 7 control-treated and n = 8 anti-HER2/CD3 TDB-treated mice) ANG2 levels as well as tissue ANG2 levels in the liver (J, n = 8 mice/treatment) and tumor (K, n = 8 mice/treatment) 24 h post treatment. Data information: Black = control-treated mice. Red = anti-HER2/CD3 TDB-treated mice. Mann–Whitney test was run to determine P-values for experiments with two groups unless otherwise indicated. Data are shown as means ± SEM unless otherwise indicated. EC, endothelial cell. Source data are available online for this figure.
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