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: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3

doi: 10.1152/ajpheart.00243.2014

Figure Lengend Snippet: A: expression of histone deacetylases (HDAC1) in both the nuclear (lanes 1 and 2) and cytosolic fractions (lanes 3 and 4) of bovine aortic endothelial cells (BAECs). B: HDAC1 and nitric oxide synthase 3 (NOS3) interact basally in BAECs shown by anti-NOS3 immunoprecipitation (IP) with immunoblot (IB) for HDAC1 from total BAEC lysate, N = 3. C: NOS3 and HDAC1 interact in COS7 cell-reconstituted system with overexpressed NOS3 and/or HDAC1-Flag tag. L, molecular mass ladder.

Article Snippet: Primary bovine aortic endothelial cells (BAECs) were purchased from Cell Applications (San Diego, CA), and COS7 (with no endogenous NOS3) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Immunoprecipitation, Western Blot, FLAG-tag

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3

doi: 10.1152/ajpheart.00243.2014

Figure Lengend Snippet: A: HDAC1-transfected cells expressed more HDAC1 protein than untransfected or vector-transfected BAECs. B: IP of anti-acetylated lysine with IB of anti-NOS3 revealed that overexpression of HDAC1 (lanes 4–6) decreased NOS3 acetylation (Ace-NOS3) with quantification by densitometry displayed in C. N = 3; *P < 0.05. D: overexpression of HDAC1 in BAECs did not significantly affect total NOS3 expression or NOS3 phosphorylation status at sites T497, S635, S1179 with quantification by densitometry displayed in E. N = 3; P > 0.05.

Article Snippet: Primary bovine aortic endothelial cells (BAECs) were purchased from Cell Applications (San Diego, CA), and COS7 (with no endogenous NOS3) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Transfection, Plasmid Preparation, Over Expression, Expressing, Phospho-proteomics

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3

doi: 10.1152/ajpheart.00243.2014

Figure Lengend Snippet: Overexpression of HDAC1 in BAECs reduced nitrite production under basal and endothelin-1 (ET-1; 100 nM, 1 h)-stimulated states (A), whereas ionomycin (3 μM, 1 h)-stimulated nitrite production was not significantly affected by overexpression of HDAC1 (B). Ionomycin did not significantly affect NOS3 acetylation (C), but ionomycin significantly reduced NOS3 T497 phosphorylation (D) after 5 or 60 min of treatment (E). N = 3 to 4. *P < 0.05 compared with vector + vehicle; †P < 0.05 compared with respective vehicle treatment. AU, arbitrary units.

Article Snippet: Primary bovine aortic endothelial cells (BAECs) were purchased from Cell Applications (San Diego, CA), and COS7 (with no endogenous NOS3) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Over Expression, Phospho-proteomics, Plasmid Preparation

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3

doi: 10.1152/ajpheart.00243.2014

Figure Lengend Snippet: HDAC1 small-interfering RNA (siRNA) knockdown in BAECs resulted in a 50% reduction in HDAC1 protein expression (A) compared with scramble siRNA control BAECs (B). C: HDAC1 knockdown resulted in an increase in nitrite production under basal and ET-1 (100 nM, 1 h)-stimulated states, whereas ionomycin (3 μM, 1 h)-stimulated nitrite production was not significantly affected by HDAC1 knockdown. D: NOS3 acetylation was similar between scramble and HDAC1 siRNA cells as quantified in E. F: NOS3 phosphorylation levels were not significantly different from scramble controls. G: densitometry of the phosphorylation sites of NOS3. N = 3–5. *P < 0.05 compared with scramble + vehicle; †P < 0.05 compared with respective vehicle treatment.

Article Snippet: Primary bovine aortic endothelial cells (BAECs) were purchased from Cell Applications (San Diego, CA), and COS7 (with no endogenous NOS3) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Small Interfering RNA, Knockdown, Expressing, Control, Phospho-proteomics