14-0627-80

Chidiac R, Abedin M, Macleod G, Yang A, Thibeault PE, Blazer LL, Adams JJ, Zhang L, Roehrich H, Jo HN, Seshagiri S, Sidhu SS, Junge HJ, Angers S
EMBO molecular medicine 2021 Jul 7;13(7):e13977

Colonization of dermal arterioles by Neisseria meningitidis provides a safe haven from neutrophils.

Manriquez V, Nivoit P, Urbina T, Echenique-Rivera H, Melican K, Fernandez-Gerlinger MP, Flamant P, Schmitt T, Bruneval P, Obino D, Duménil G
Nature communications 2021 Jul 27;12(1):4547

Using CRISPR-Cas9 to quantify the contributions of O-glycans, N-glycans and Glycosphingolipids to human leukocyte-endothelium adhesion.

Stolfa G, Mondal N, Zhu Y, Yu X, Buffone A Jr, Neelamegham S
Scientific reports 2016 Jul 26;6:30392

Effect of nicotine and porphyromonas gingivalis lipopolysaccharide on endothelial cells in vitro.

An N, Andrukhov O, Tang Y, Falkensammer F, Bantleon HP, Ouyang X, Rausch-Fan X
PloS one 2014;9(5):e96942

Endothelial leukocyte adhesion molecule 1: an inducible receptor for neutrophils related to complement regulatory proteins and lectins.

Bevilacqua MP, Stengelin S, Gimbrone MA Jr, Seed B
Science (New York, N.Y.) 1989 Mar 3;243(4895):1160-5

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-CD62E (E-selectin) Monoclonal Antibody (Product # 14-0627-82) and 67, 115 kDa bands corresponding to CD62E were observed across the tissues tested. Tissue extracts (30 µg lysate) of Mouse Liver (Lane 1), Mouse Spleen (Lane 2) and Mouse White Adipose (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Staining of TNF alpha-stimulated Human Umbilical Vein Endothelial Cells (HUVEC) with 0.5 µg of Mouse IgG1 kappa Isotype Control Purified (Product # 14-4714-82) (open histogram) or 0.5 µg of Anti-Human CD62E (E- Selectin) Purified (filled histogram) followed by F (ab')2 Anti-Mouse IgG PE (Product # 12-4012). Total viable cells were used for analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Effect of nicotine on the P. gingivalis LPS-induced protein expression of pro-inflammatory mediators in HUVECs. HUVECs were stimulated by P. gingivalis LPS in the presence or absence of nicotine (10 uM-10 mM) for 4, 24, and 72 h. After stimulation, the surface expression levels of ICAM-1 (A), VCAM-1 (B), and E-selectin (C) were measured by flow cytometry, and the quantity of MCP-1 (D) and IL-8 (E) in conditioned media was measured by ELISA. Each value represents mean +-SD of three independent assays. Non-stimulated HUVECs were used as a control. The protein expression levels of pro-inflammatory mediators were not analyzed after stimulation with 10-mM nicotine for 24 and 72 h because the cells were not viable. * - significantly different between groups, p