Fig 1: TMAs and western blot validation analysis of AKAP12, DCBLD2, NT5E and SPON1. a. “Columns from left to right” indicate immunostaining pattern in normal colonic samples and CRC cores negative and positive for each marker AKAP12, DCBLD2, NT5E, SPON1. Black arrows indicate immunohistochemical staining pattern in normal or malignant colonic cells. White arrows indicate the immunostaining distribution in the stromal compartment (endothelial, regulatory T-cells and macrophages) characteristic of NT5E expression pattern. Magnification 10X; b. Number of positive cases detected on TMA validation series comprising tumor specimens (TUM) and a subgroup of matched normal colonic mucosa (NM). Error bars indicate the standard deviation from the mean (p < 0.05); c. Four representative frozen CRC specimens (T) and matched normal mucosa (N) were identified in the same cohort of patients and analyzed by immunoblot. Molecular weight markers are indicated in kilodaltons. ß-tubulin was used as loading control to normalize band intensities.
Fig 2: Cross-talk between TNF-α and PPARγ signaling regulates DCBLD2 and NT5E intracellular levels in CRC cell lines. a. Western blotting analysis shows PPARγ expression in HT29 and RKO CRC derived cell lines, as a model to investigate variations of NTE5 levels in response to TNF-α treatment. b. PPARγ-dependent protein induction of NT5E by TNF-α in CRC cells treated with 12,5 ng/ml TNF-α at different time points and analyzed by western blotting. c and d. Time course of NT5E and DLBLC2 mRNA and protein regulation by troglitazone (TGZ), a PPARγ agonist, in the same cells. β-actin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as control to normalize expression levels in Western blotting and Real-time RT-PCR analysis. The results are expressed as means ±S.D. of three independent experiments. * p < 0.05, **p < 0.01.
Fig 3: Survival curves of our cohort as function of the selected genes. a. Survival curves on our CRC validation series, categorized as having high (red curve) and low (green curve) AKAP12, DCBLD2 and NT5E expression. p-value for the null hypothesis of equal population survival curves is provided by log-rank test in each graph; b. Survival curves estimated combining the expression (high and low) of all 3 markers (AKAP12, DLBLC2, NTE5). Red curve represents the combination high/high; the blue curve represents the combination high/low; the black curve represents the combination low/high; the green curve represents the combination low/low.
Supplier Page from Abcam for Anti-DCBLD2/ESDN antibody