Journal: bioRxiv

Article Title: Intraprocedural endothelial cell seeding of arterial stents via biotin/avidin targeting mitigates in-stent restenosis

doi: 10.1101/2022.05.25.493423

Figure Lengend Snippet: Step 1: Metal samples are exposed to aqueous polyallylamine bisphosphonate (PAB) solution resulting in permanent surface modification via formation of coordination bonds between the metal atoms and bisphosphonate groups of PAB to exhibit primary amine groups available for chemical conjugation reactions. Step 2: Aminated metal substrate is reacted with an amine-reactive biotinylation agent (sulfo-NHS-LC-LC-biotin). Step 3: Biotinylated metal substrate is exposed to avidin to attain a monolayer of avidin on the metal surface. Step 4: Endothelial cells are surface modified with biotin via the reaction of sulfo-NHS-LC-LC-biotin with amine group of proteins on the cell surface. Step 5: Biotinylated endothelial cells are brought in contact with avidin-modified metal samples resulting in augmented cell binding to the metal substrate (Step 6).

Article Snippet: Primary rat aortic endothelial cells (RAEC; Cell Applications, San Diego, CA) were cultured in EGM2 medium (Lonza, Walkersville, MD) and used at passages 4-7.

Techniques: Modification, Conjugation Assay, Avidin-Biotin Assay, Binding Assay

Journal: Journal of Cell Science

Article Title: Contribution of protein–protein interactions to the endothelial-barrier-stabilizing function of KRIT1

doi: 10.1242/jcs.258816

Figure Lengend Snippet: Membrane-targeted KRIT1-RE does not rescue endothelial barrier function. (A) Permeability of transduced monolayers to 40 kDa FITC-dextran. Data shown are mean permeability±s.e.m., normalized to scramble shRNA alone (scr), from n =5 independent experiments. (B) TEER of confluent HPAEC monolayers. Resistance reading of an empty FN-coated Transwell was subtracted from resistance values of Transwells containing HPAEC, then multiplied by the growth area of the wells, yielding Ω*cm 2 values. Data shown are mean Ω*cm 2 values±s.e.m., normalized to scramble shRNA alone (negative control). * P <0.05 by Tukey post-hoc testing vs scramble shRNA alone. # P <0.05 by Tukey post-hoc testing vs shKRIT1 alone. P <0.0001 by one-way ANOVA.

Article Snippet: Human pulmonary artery endothelial cells (HPAEC; Cell Applications, Inc., San Diego, CA, Lot #2228) were cultured in Dulbecco's Modified Eagle's Medium DMEM/F-12 (1:1 ratio), supplemented with 5% fetal bovine serum (FBS), 1% antibiotic-antimycotic solution (Gibco/Thermo Scientific, Waltham, MA), 1% endothelial cell growth supplement (ECGS; ScienCell, Carlsbad, CA), and 50 μM heparin (Calbiochem, La Jolla, CA).

Techniques: Membrane, Permeability, shRNA, Negative Control