Figure 4 from Collagen XVII Promotes Pancreatic Ductal Adenocarcinoma Tumor Growth through Regulation of PIK3R5

Viability and metastatic potential of PDAC cells is dependent on collagen XVII. A, Western blot analysis of collagen XVII in MGH1319, MGH1275, and MGH1108 cells expressing an NTC shRNA and COL17A1-specific shRNAs. The expression of β-actin served as a control. B, Resazurin-based cell viability assays were performed using MGH1319, MGH1275, and MGH1108 cells expressing NTC and COL17A1-specific shRNAs. The mean (±SEM) cell viability relative to NTC-expressing cells of three independent experiments performed in triplicate is shown. ***, P post hoc test). C, Representative scratch areas at 0 and 24 hours of MGH1319, MGH1275, and MGH1108 cells expressing NTC and COL17A1-specific shRNAs. D, The mean (±SEM) scratch closure area relative to 0 hours from three independent biological experiments is shown for MGH1319, MGH1275, and MGH1108. ***, P P post hoc test). E, The mean (±SEM) migration of MGH1319 cells expressing COL17A1-specific shRNAs relative to cells expressing NTC, as measured by a transwell migration assay. Three independent infections of MGH1319 are shown. *, P P post hoc test). F, Representative images of COX IV IHC staining of lungs harvested from mice 4 weeks after injection with MGH1319 cells expressing an NTC vector or COL17A1-specific shRNAs. Scale bars, 100 µm. G, Percentage of COX IV–positive area (±SEM) in the lungs of mice with metastasis formed by MGH1319 cells expressing either an NTC or COL17A1-specific shRNAs (n = 6 for each shRNA). ****, P post hoc test).