| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-E-Selectin (CD62E) antibody, Mouse monoclonal | Sigma Aldrich (UK) | Cat# S9555; RRID: AB_1078476 |
| Chicken anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 | ThermoFisher Scientific (USA) | Cat# A-21200; RRID: AB_2535786 |
| Anti-Mouse IgG (whole molecule)−Alkaline Phosphatase antibody produced in goat | Sigma Aldrich (UK) | Cat# A9316; RRID: AB_258446 |
| Anti-HiBiT antibody, mouse monoclonal | Promega (USA) | Cat# N7200; RRID: AB_2924793 |
| Anti-α-tubulin antibody, rabbit monoclonal | Cell Signalling Technologies (The Netherlands) | Cat# 2144; RRID: AB_2210548 |
| IRDye 800CW goat anti-mouse IgG secondary antibody (H+L) | Li-Cor Biosciences (UK) | Cat# 925-32210; RRID: AB_2687825 |
| IRDye 680RD goat anti-rabbit IgG secondary antibody (H+L) | Li-Cor Biosciences (UK) | Cat# 925-68071; RRID: AB_2721181 |
| Chemicals, peptides, recombinant proteins | ||
| FuGENE HD Transfection Reagent | Promega (USA) | Cat# E2311 |
| Opti-MEM reduced serum medium | ThermoFisher Scientific (USA) | Cat# 11058021 |
| Formalin solution neutral buffered, 10% | Sigma Aldrich (UK) | Cat# HT501128 |
| Chicken serum | Sigma Aldrich (UK) | Cat# C5405 |
| Goat serum | Abcam (UK) | Cat# ab7481 |
| Bovine serum albumin (BSA) | Sigma Aldrich (UK) | Cat# 03117332001 |
| Glycine | Sigma Aldrich (UK) | Cat# G8898 |
| bisBenzimide H33342 trihydrochloride | Sigma Aldrich (UK) | Cat# B2261 |
| Human Large Vessel Endothelial Cell Basal Medium (Medium 200) | ThermoFisher Scientific (Gibco) (USA) | Cat# M200500 |
| Large Vessel Endothelial Supplement (LVES) | ThermoFisher Scientific (Gibco) (USA) | Cat# A1460801 |
| Dulbecco’s Modified Eagle’s Medium (DMEM) | Sigma Aldrich (UK) | Cat# D6429 |
| Fetal Calf Serum (FCS) | Sigma Aldrich (UK) | Cat# F2242 |
| Trypsin-EDTA Solution x10 | Sigma Aldrich (UK) | Cat# T4174 |
| Dulbecco's Phosphate Buffered Saline | Sigma Aldrich (UK) | Cat# D8537 |
| Poly-D-Lysine hydrobromide | Sigma Aldrich (UK) | Cat# P6407 |
| Gelatin solution | Sigma Aldrich (UK) | Cat# G1393 |
| LB Broth (Lennox) | Sigma Aldrich (UK) | Cat# L3022 |
| Tumor Necrosis Factor-α human | Sigma Aldrich (UK) | Cat# H8916 |
| Recombinant Human VEGF 165 Protein (with carrier) | R&D systems (USA) | Cat# 293-VE-050 |
| Histamine dihydrochloride | Sigma Aldrich (UK) | Cat# H7250 |
| IL-1 alpha human | Sigma Aldrich (UK) | Cat# SRP6295 |
| IL-1 beta human | Sigma Aldrich (UK) | Cat# SRP3083 |
| Lipopolysaccharides from Escherichia coli O55:B5 | Sigma Aldrich (UK) | Cat# L2880 |
| RIPA buffer | Sigma Aldrich | Cat# R0278 |
| Roche cOmplete, mini protease inhibitor cocktail | Sigma Aldrich | Cat# 11836153001 |
| Pierce™ BCA Protein Assay Kit | ThermoFisher (UK) | Cat# 23227 |
| Invitrogen NuPAGE™ LDS Sample Buffer | ThermoFisher (UK) | Cat# NP0007 |
| Invitrogen NuPAGE™ Sample Reducing Agent | ThermoFisher (UK) | Cat# NP0004 |
| Invitrogen NuPAGE™ MES SDS Running Buffer | ThermoFisher (UK) | Cat# NP0002 |
| Invitrogen Nitrocellulose/Filter Paper Sandwich, 0.45 μm, 8.3 x 7.3 cm | ThermoFisher (UK) | Cat# LC2001 |
| Invitrogen NuPAGE™ Transfer Buffer | ThermoFisher (UK) | Cat# NP00061 |
| Intercept (TBS) blocking buffer | Li-Cor Biosiences (UK) | Cat# 927-60001 |
| Li-CoR Total Protein Stain | Li-Cor Biosiences (UK) | Cat# 926-11010 |
| GeltrexTM LDEV-free reduced growth factor basement membrane matrix | ThermoFisher Scientific (USA) | Cat# A1413202 |
| Purified Cas 9 | IDT (USA) | Cat# 1081059 |
| Electroporation enhancer | IDT (USA) | Cat# 1075916 |
| Ingenio® Electroporation Solution | Mirus Bio (USA) | Cat# MIR 50114 |
| Critical commercial assays | ||
| Nano-Glo luciferase assay (Furimazine) | Promega (USA) | Cat# N1130 |
| Nano-Glo HiBiT Lytic Detection System | Promega (USA) | Cat# N3040 |
| Experimental models: Cell lines | ||
| Human umbilical vein endothelial cells (HUVECs) (newborn male single donor) | ThermoFisher Scientific (USA) | Cat# C0035C Lot#: 1606186 |
| Human telomerase reverse transcriptase 2 (hTERT2) HUVECs (female) | ATCC (Virginia, USA) | Cat# CRL-4053 Lot# 70023166 |
| Oligonucleotides | ||
| E-selectin guide DNA | IDT (USA) | Custom |
| TRACR RNA | IDT (USA) | Cat# 1073191 |
| HiBiT E-selectin Repair DNA | IDT (USA) | Custom |
| Recombinant DNA | ||
| E-selectin vector | Vector Builder (USA) | Custom |
| Software and algorithms | ||
| GraphPad Prism 7.02 | GraphPad Software | www.graphpad.com |
| Zen 2012 | Zeiss | www.zeiss.com |
| ImageJ Fiji 1.53 | National Institute of Health | www.fiji.sc |
| Other | ||
| μ-Slide 8 well plate - ibidiTreat: #1.5 polymer coverslip, tissue culture treated, sterilized | Thistle Scientific (Ibidi) | Cat# 80826 |
| Greiner 96 well plate (White) | Greiner Bio-One | Cat# 655089 |
| Greiner 96 well plate (Black) | Greiner Bio-One | Cat# 655090 |
| Corning® 96-well Clear Flat Bottom Polystyrene TC-treated Microplate | Corning (USA) | Cat# 3596 |
| 4-chamber 35mm dish | Cellvis (USA) | Cat# D35C4-20-1.5-N |
| Fisherbrand™ Electroporation Cuvettes Plus™ | Fisher scientific | Cat# FB102 |
| Nucleofector® 2b Device | Lonza | Cat# AAB-1001 |
| E.Z.N.A.® Tissue DNA Kit | Omega Bio-tek (USA) | Cat# D3396-01 |
| Wizard SV Gel and PCR Clean-Up System | Promega (USA) | Cat# A9281 |
| BamHI enzyme | Promega (USA) | Cat# R6021 |
| XhoI enzyme | Promega (USA) | Cat# R6161 |
| DH5 alpha Chemically Competent cells | invitrogen | Cat# 18265-017 |
| T4 DNA ligase | New England Biolabs | Cat# MO202S |
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