Journal: Oncotarget

Article Title: YBX1/YB-1 induces partial EMT and tumourigenicity through secretion of angiogenic factors into the extracellular microenvironment

doi:

Figure Lengend Snippet: a. Cell migration over 24 hr assessed by wound-healing assays and phase-contrast microscopy (4 × magnification). b. Cell migration as assessed by transwell assay using 8.0 μm membrane culture inserts. 24 h after cell seeding, migrating cells that passed through the transwell were stained with DAPI and counted ( n = 3; average ± SEM). c. Transwell invasion assays were conducted using 8.0 μm membrane inserts coated with 1 mg/mL Matrigel. 24 h after cell seeding, invading cells were stained with DAPI and counted ( n = 3; average ± SEM). d. Colony forming assays were performed in soft agar using CytoSelect 96-well cell transformation kit. 2,500 cells were inoculated and cultured for 7 days. Number of colonies were manually counted from 5 random fields of view (for each replicate) and colony size measured and quantified. Images are representative. (scale bar: 100 μm) ( n = 3; average ± SEM; ** P < 0.01).

Article Snippet: Cells were overlaid onto Transwell ® polycarbonate membrane cell culture inserts (8.0 μm pore size, Corning), and inserts placed into 24-well companion plates.

Techniques: Migration, Microscopy, Transwell Assay, Membrane, Staining, Transformation Assay, Cell Culture

Journal: Oncotarget

Article Title: YBX1/YB-1 induces partial EMT and tumourigenicity through secretion of angiogenic factors into the extracellular microenvironment

doi:

Figure Lengend Snippet: a. 5 × 10 4 2F-2B cells were conditioned with 30 μg of MDCK, MDCK YBX1 or 21D1 cell-derived secretome. Cell migration over 24 hr was assessed using Transwell inserts (8.0 μm pore size). Migrating cell nuclei were stained with DAPI, imaged and quantified ( n = 3; average ± SEM; ** P < 0.01). b. Cell-derived secretome was profiled using antibody-based angiogenesis arrays. Expression of 55 target proteins in the secretome was assessed. c. Relative expression of angiogenic modulators in cell-derived secretome ( n = 3; average intensity ± STD. * P < 0.05, ** P < 0.01).

Article Snippet: Cells were overlaid onto Transwell ® polycarbonate membrane cell culture inserts (8.0 μm pore size, Corning), and inserts placed into 24-well companion plates.

Techniques: Derivative Assay, Migration, Pore Size, Staining, Expressing

Journal: PLoS ONE

Article Title: Streptococcus pneumoniae potently induces cell death in mesothelial cells

doi: 10.1371/journal.pone.0201530

Figure Lengend Snippet: A) MeT-5A cells were incubated with cell-free bacteria conditioned media. B) S . pneumoniae was loaded into the top chamber of a Transwell® insert and viability of MeT-5A cells was determined in the bottom chamber. Cell viability was assessed 24 hr post-treatment. * Denotes significantly higher than vehicle control.

Article Snippet: Transwell experiments were performed where MeT-5A cells were cultured in 24-well plates and 200 μL of a S . pneumoniae suspension (2 x 10 7 CFU/mL) was transferred into the upper chamber of Corning® Transwell® polyester membrane cell culture inserts (6.5 mm diameter, 0.4 μm pore size; Sigma-Aldrich).

Techniques: Incubation

Journal: PLoS ONE

Article Title: Intraperitoneal infusion of mesenchymal stem cell attenuates severity of collagen antibody induced arthritis

doi: 10.1371/journal.pone.0198740

Figure Lengend Snippet: (A) Expression of multiple chemokines in peritoneal mononuclear cells of CAIA mice treated with or without MSCs. Mononuclear cells were isolated from peritoneal wash fluid from CAIA mice. (B) In vitro scratch assay to examine MSC migration in the absence or presence of SDF-1α and RANTES. The number of MSCs migrating into the gap was counted every 2 h (between 0 and 12 h) by three independent examiners. Representative images taken at 0 and 12 h are shown. (C) Transwell migration assay to examine MSC migration in response to SDF-1α and RANTES. The two chambers were separated by a cell permeable polycarbonate membrane. The upper chamber was loaded with MSCs and the lower chamber was filled with DMEM containing bovine serum albumin (control), SDF-1α, or RANTES. After 7 h, the number of MSCs migrating into the lower chamber was counted by three independent observers. Data are expressed as the mean ± standard error of the mean (SEM). ** p < 0.01; *** p < 0.001.

Article Snippet: Chemotaxis of MSCs was evaluated using commercially available Transwell® polycarbonate membrane cell culture inserts in 24-well plates (CLS3422, Sigma-Aldrich, St. Louis, MO, USA) [ ].

Techniques: Expressing, Isolation, In Vitro, Wound Healing Assay, Migration, Transwell Migration Assay