Journal: Arthritis Research & Therapy

Article Title: Fucosyltransferase 1 mediates angiogenesis, cell adhesion and rheumatoid arthritis synovial tissue fibroblast proliferation

doi: 10.1186/ar4456

Figure Lengend Snippet: α(1,2)-linked fucosylated proteins are expressed in rheumatoid arthritis (RA). (A) RA synovial tissue (ST) homogenates contained more α(1,2)-linked fucosylated proteins than did osteoarthritis (OA) or normal (NL) STs (normalized to total protein concentration). Results are expressed as a ratio of the amount of fucosylated BSA/total proteins in the ST homogenates using BSA as a standard. (B) Photographs of RA ST. The far left panel shows staining with mouse anti-collagen 1. The middle panel shows staining with Ulex Europeaus Agglutinin 1 lectin (UEA-1) and goat anti-UEA-1. The right panel shows merging of the left panel and middle panel. Yellow indicates α(1,2)-linked fucosylated proteins associated with RA ST fibroblasts and the blue indicates DAPI staining of the tissue (original magnification 100×). (C) α(1,2)-linked fucosylated proteins in RA synovial fibroblast-conditioned medium were significantly higher than in OA and NL synovial fibroblast-conditioned medium. (D) α(1,2)-linked fucosylated proteins in RA synovial fibroblast cell lysates were significantly higher than in NL synovial fibroblast cultures: 2′fucosyllactose-bovine serum albumin (2′FL-BSA) was used as a standard.

Article Snippet: To determine if α(1,2)-linked proteins were expressed on RA ST synovial fibroblasts, mouse anti-human collagen-1 (Abcam, Cambridge, MA, USA) and goat anti-UEA-1 (Vector laboratories Inc.) were used.

Techniques: Protein Concentration, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Dynamic Endothelial Stalk Cell–Matrix Interactions Regulate Angiogenic Sprout Diameter

doi: 10.3389/fbioe.2021.620128

Figure Lengend Snippet: Endothelial stalk cell proliferation is positively correlated with sprout diameter. (A) Representative images (max intensity projections) of invading endothelial cells (ECs) in response to varying phorbol 12-myristate 13-acetate (PMA). All conditions were cultured for 5 days with 250 nM sphingosine 1-phosphate (S1P) in endothelial cell growth medium 2 (EGM2; supplemented with indicated PMA) added to the chemokine channel within 3 mg ml –1 collagen hydrogels. Ulex Europaeus Agglutinin-1 (UEA; white), nuclei (magenta), 5-ethynyl-2′-deoxyuridine (EdU; cyan), and yellow dashed lines indicate parent vessel edge. (B,C) Quantifications of proliferation and sprout diameter as a function of PMA. For proliferation: n ≥ 12 per condition and for sprout diameter: n ≥ 16 per condition. (D) Relationship between proliferation and sprout diameter, with red dashed line indicating a linear regression and statistical analysis performed by Pearson’s correlation. Sample size for each mean is identical to those of panels (B,C) . All data presented as mean ± SD; *indicates a statistically significant comparison with P < 0.05 (one-way ANOVA).

Article Snippet: DyLight 649 labeled Ulex Europaeus Agglutinin-1 (UEA, 1:200, Vector Labs, Burlingame, CA, United States) was utilized to visualize EC morphology in samples stained with EdU due to the incompatibility of EdU ClickIT chemistry with phalloidin staining.

Techniques: Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Dynamic Endothelial Stalk Cell–Matrix Interactions Regulate Angiogenic Sprout Diameter

doi: 10.3389/fbioe.2021.620128

Figure Lengend Snippet: Denser matrix that slows sprout invasion leads to smaller sprout diameters. (A) Representative images (max intensity projections) of invading endothelial cells (ECs) in response to varying collagen density. All conditions were cultured for 3 days with 250 nM sphingosine 1-phosphate (S1P) in endothelial cell growth medium 2 [EGM2; supplemented with 25 ng ml –1 phorbol 12-myristate 13-acetate (PMA)] added to the chemokine channel within collagen hydrogels of the indicated density. Nuclei (magenta), 5-ethynyl-2′-deoxyuridine (EdU; cyan), Ulex Europaeus Agglutinin-1 (UEA; white), and yellow dashed lines indicate parent vessel edge. (B–D) Quantifications of proliferation, invasion speed, and sprout diameter as a function of collagen hydrogel density. For proliferation: n ≥ 6 per condition, for invasion speed: n ≥ 60 per condition, and for sprout diameter: n ≥ 100 per condition. (E,F) Relationships between proliferation and sprout diameter (E) and invasion speed and sprout diameter (F) , with red dashed lines indicating a linear regression and statistical analyses performed by Pearson’s correlation. Sample size for each mean is identical to those of panels (B–D) . All data presented as mean ± SD; *indicates a statistically significant comparison with P < 0.05 (one-way ANOVA).

Article Snippet: DyLight 649 labeled Ulex Europaeus Agglutinin-1 (UEA, 1:200, Vector Labs, Burlingame, CA, United States) was utilized to visualize EC morphology in samples stained with EdU due to the incompatibility of EdU ClickIT chemistry with phalloidin staining.

Techniques: Cell Culture