Journal: International Journal of Cancer

Article Title: Transcriptional profiling of breast cancer‐associated lymphatic vessels reveals VCAM‐1 as regulator of lymphatic invasion and permeability

doi: 10.1002/ijc.32594

Figure Lengend Snippet: VCAM‐1 blockade reduces lymphatic permeability. ( a , b ) Representative FACS histograms (left panels) and staining quantification (right panels) of the classic VCAM‐1 receptors integrin α4 (Itga4), integrin α9 (Itga9) and integrin β1 (Itgb1) in 4T1‐1F8 cells ( a ) and cultured mouse LECs with or without TNF‐α treatment for 24 hr ( b ; n = 3). ( c ) Adhesion of fluorescently labeled 4T1‐1F8 cells to monolayers of cultured mouse LECs pretreated with TNF‐α or not and in the presence of control IgG or VCAM‐1 blocking antibodies ( n = 3 wells/condition; 1 representative of 6 individual experiments is shown). ( d ) In vitro permeability of mouse LEC monolayers grown on transwell inserts (0.4 μm pore size) toward 70 kD FITC‐dextran. LEC monolayers were pretreated with TNF‐α overnight and subsequently incubated with control or 4T1 CM in presence of control IgG or anti‐VCAM‐1 antibodies ( n = 3 wells/condition; one representative of three individual experiments is shown). Statistical test refers to 4T1‐1F8 CM—rat IgG1 compared to 4T1‐1F8 CM—anti‐VCAM‐1. ( e ) Representative images (left panels) and quantification (right panel) of VE‐cadherin staining in mouse LEC monolayers that were pretreated with TNF‐α overnight and subsequently incubated with control or 4T1 CM in presence of control IgG or anti‐VCAM‐1 antibodies ( n = 10 random images/condition; * p < 0.05). [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: Mouse LECs were seeded on fibronectin/collagen‐coated transwell inserts (0.4 μm pore size, Costar, Washington, DC) and starved O/N in DMEM/F12 + 1% FBS with or without TNF‐α (40 ng/ml, Peprotech).

Techniques: Permeability, Staining, Cell Culture, Labeling, Blocking Assay, In Vitro, Incubation

Journal: Fluids and barriers of the CNS

Article Title: Subcellular trafficking and transcytosis efficacy of different receptor types for therapeutic antibody delivery at the blood‒brain barrier.

doi: 10.1186/s12987-023-00480-x

Figure Lengend Snippet: Fig. 2 Characterization of receptor expression in two human cell barrier models. A Sorting receptors TfR and sortilin are both expressed in hiPSC-BECs and Caco-2 cells and localized at intracellular surfaces. The polarized receptors podocalyxin and CD133 are both expressed in the apical surface facing the luminal side of the Transwell chamber. B Micrographs showing maximum projected z-stacks with side view zx images above and xy images below. Cells are stained for ZO1 in white and the indicated receptors in green. Scale bars are 10 µm. See also Additional file 2: Fig S2

Article Snippet: Then, 500,000 cells were seeded in EC medium onto collagen IV/fibronectin-coated Transwell thinserts (12- well format, 1.13 cm2, GREINER BIO-ONE, Cat. No.: GR-665641) with or without astrocyte noncontact coculture.

Techniques: Expressing, Staining

Journal: Fluids and barriers of the CNS

Article Title: Subcellular trafficking and transcytosis efficacy of different receptor types for therapeutic antibody delivery at the blood‒brain barrier.

doi: 10.1186/s12987-023-00480-x

Figure Lengend Snippet: Fig. 4 Transcytosis of cargo antibodies through cell barriers. A ELISA measurement of cargo antibody in the Transwell bottom acceptor chamber in relation to the added antibody in the top donor chamber given as transcytosed IgG percentage through the hiPSC-BEC layer. The red stippled line indicates the lower level of detection (approx. 50 pM) for the ELISA setup. B ELISA quantification of cargo antibody transcytosis through the Caco-2 cell layer. Statistical significance was determined using two-way ANOVA with Tukey´s multiple comparisons test based on four independent experiments. See also Additional file 2: Fig S5

Article Snippet: Then, 500,000 cells were seeded in EC medium onto collagen IV/fibronectin-coated Transwell thinserts (12- well format, 1.13 cm2, GREINER BIO-ONE, Cat. No.: GR-665641) with or without astrocyte noncontact coculture.

Techniques: Enzyme-linked Immunosorbent Assay