Journal: Oncology Reports

Article Title: Combination treatment with VPA and MSCs-TRAIL could increase anti-tumor effects against intracranial glioma

doi: 10.3892/or.2021.7937

Figure Lengend Snippet: Increased migration ability of MSCs-TRAIL toward VPA-treated tumors and VPA-induced increase of CXCR4 expression in MSCs-TRAIL. (A) Upregulation of CXCR4 expression was identified via western blot analysis following addition of VPA to MSCs and MSCs-TRAIL culture medium. β-actin was used as a loading control. (B) Effect of treatment with SDF-1 and AMD3100 on the migration of MSCs-TRAIL was examined using a Transwell migration assay. Magnification, ×100. Data are presented as the mean ± SD. *P<0.05 and **P<0.01 vs. the treatment with SDF-1 or VPA alone (one-way ANOVA with Bonferroni multiple comparison test). (C) Migratory ability of MSCs-TRAIL in response to conditioned medium from untreated or VPA-treated tumors was determined using a Transwell plate (8-µm pores). Representative photomicrographs of stained filters show migrated cells. Magnification, ×100. Data are presented as the mean ± SD. **P<0.01 vs. treatment of MSCs-TRAIL or VPA (one-way ANOVA with Bonferroni multiple comparison test). (D) To measure the in vivo migration capacity of MSCs-TRAIL, PKH67(green)-labeled MSCs-TRAIL were injected together with PKH26(red)-labeled VPA-treated MSCs-TRAIL into the contralateral hemisphere of tumors in a glioma model. Nuclei were stained with DAPI (blue). Box a and b, Tumor area; Box N, Normal area; T, tumor area. Visualization was conducted using a confocal microscope (magnification, ×100). Dotted line, tumor edge. VPA, valproic acid; TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; DR, death receptor; MSCs-TRAIL, tumor necrosis factor-related apoptosis-inducing ligand-secreting human bone marrow-derived mesenchymal stem cells; SDF-1, stromal cell-derived factor-1; CXCR4, C-X-C chemokine receptor type 4; CM, conditioned medium; SFM, serum free medium.

Article Snippet: The migratory abilities of MSCs and MSCs-TRAIL were determined using Transwell plates (Costar; Corning, Inc.) that were 6.5-mm in diameter with 8-μm pore filters.

Techniques: Migration, Expressing, Western Blot, Control, Transwell Migration Assay, Comparison, Staining, In Vivo, Labeling, Injection, Microscopy, Derivative Assay

Journal: The EMBO Journal

Article Title: Endoglin mediates fibronectin/?5?1 integrin and TGF-? pathway crosstalk in endothelial cells

doi: 10.1038/emboj.2012.246

Figure Lengend Snippet: Fibronectin/integrin α5β1 switch TGF-β from a migration promoter to migration suppressor. (A) HMEC-1 were plated in the transwells coated with non-ECM, 20 μg/ml fibronectin or collagen, pretreated with or without 100 pM TGF-β1 and assessed for migration after 8 h. Quantitated data from three independent experiments ±s.d. are presented. (B) MEEC+/+ pretreated with or without 10 μg/ml α5β1 function-blocking antibody were plated in transwells coated with 20 μg/ml fibronectin, treated with or without 100 pM TGF-β1 and assessed for migration after 8 h. Quantitated data from two independent experiments ±s.d. are presented. (C) MEEC+/+ or MEEC−/− were plated in transwells coated with 20 μg/ml fibronectin, pretreated with or without 100 pM TGF-β1 and assessed for migration after 8 h. Quantitated data from three independent experiments ±s.d. are presented. (D) Wild HMEC-1 or HMEC-1 adenovirally infected with shRNA to endoglin were cultured on the Matrigel mixed with or without 50 μg/ml fibronectin with 100 pM TGF-β1 treatment for indicated times. Images were taken at 4 × microscopy, and tubules were counted using Image J. Quantitated data ±s.d. of one representative experiment out of two independent experiments are presented. (E) HMEC-1 or HMEC-1 adenovirally infected with shRNA to endoglin were cultured in Matrigel for 12 h, lysed and pro- and cleaved caspase-3 were detected using anti caspase-3 antibody. Figure source data can be found with the Supplementary data.

Article Snippet: Transwells (Costar Corning Inc., 8 μM polycarbonate membrane 6.5 mm insert, 24-well plate) were coated with 20 μg/ml fibronectin, laminin or collagen prior to plating.

Techniques: Migration, Blocking Assay, Infection, shRNA, Cell Culture, Microscopy