Journal: Cell Death and Differentiation

Article Title: Loss of c-REL but not NF- κ B2 prevents autoimmune disease driven by FasL mutation

doi: 10.1038/cdd.2014.168

Figure Lengend Snippet: FasLΔm/Δm mice had normal mTEC numbers with normal AIRE expression but loss of NF-κB2 caused abnormalities in the thymic stroma. (a) Representative confocal images of thymi from mice of the indicated genotypes at 6 weeks. Sections were stained for UEA-1 (green), AIRE (red) and keratin-5 (blue); n=3/genotype/time point. (b) Representative confocal images of thymi from mice of the indicated genotypes at 20 weeks as for (a). (c) Graphical representation of total thymic cellularity from mice of the indicated genotypes at 20 weeks of age; n=4/genotype. (d) Graphical depiction of flow cytometric analysis of TECs from mice of the indicated genotypes at 20 weeks of age, as defined; mTEChi (CD45− Ep-CAM+class II MHChi Ly51−), mTEClow (CD45− Ep-CAM+class II MHClow Ly51−), cTEC (CD45− Ep-CAM+class II MHC+ Ly51+); n=4/genotype. (e) Graphical depiction of flow cytometric analysis of mTEChiAIRE+ mTECS (CD45− Ep-CAM+class II MHChi Ly51− AIRE+) from mice of the indicated genotypes at 20 weeks of age. (f) Percentages of thymocytes that are CD4 single positive (CD4+ CD8−) or CD8 single positive (CD4−CD8+) from mice of the indicated genotypes at 3 months of age; n=4/genotype. *P<0.05, **P<0.005

Article Snippet: Primary Abs included biotin-conjugated anti-UEA-1 (Vector Laboratories, Burlingame, CA, USA), rabbit anti-keratin-5 (MK5, Covance, Biolegend, San Deigo, CA, USA) and Alexa 405-conjugated anti-AIRE.

Techniques: Expressing, Staining

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: UEA-1, 2 μg/ml (Vector laboratories Inc, Burlingame, CA, USA) was added for 90 minutes followed by 10 μg/ml biotinylated goat anti-UEA-1 antibody (Vector laboratories Inc) for 60 minutes, then streptavidin-horseradish peroxidase (HRP) for 30 minutes.

Techniques: Protein Concentration, Staining

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