Journal: Advanced Healthcare Materials

Article Title: Plasma‐Polymerized Nanoparticles Presenting Fibrillin‐1 Drive Rapid Re‐Endothelialization of Vascular Grafts

doi: 10.1002/adhm.202503360

Figure Lengend Snippet: PPN‐PF8 functionalization of ePTFE improves endothelial attachment in vitro. A) Representative images of SEM and F‐actin staining. B) HCAEC attachment and C) spreading on ePTFE, ePTFE with passively bound PF8 (ePTFE‐PF8), PPN‐coated ePTFE (PPN), and PPN‐PF8 functionalized ePTFE, n = 3 per sample. Scale bar = 300 µm. * p < 0.05, *** p < 0.001 versus ePTFE.

Article Snippet: Human coronary artery endothelial cells (HCAEC; Cell Applications) were cultured in MesoEndo medium (Cell Applications) at 37 °C in 5% CO 2 .

Techniques: In Vitro, Staining

Journal: Advanced Healthcare Materials

Article Title: Plasma‐Polymerized Nanoparticles Presenting Fibrillin‐1 Drive Rapid Re‐Endothelialization of Vascular Grafts

doi: 10.1002/adhm.202503360

Figure Lengend Snippet: PPN‐PF8 functionalization of ePTFE improves endothelial proliferation in vitro. A) Representative images of SEM and F‐actin staining of HCAEC proliferation on days 1 and 3. B) HCAEC proliferation and C) spreading on ePTFE, ePTFE with passively bound PF8, PPN, and PPN‐PF8 functionalized ePTFE. * p < 0.05, *** p < 0.001 versus ePTFE on each day, Scale bar = 300 µm. n = 3 per sample.

Article Snippet: Human coronary artery endothelial cells (HCAEC; Cell Applications) were cultured in MesoEndo medium (Cell Applications) at 37 °C in 5% CO 2 .

Techniques: In Vitro, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Protective role of N-acetylcysteine and Sulodexide on endothelial cells exposed on patients’ serum after SARS-CoV-2 infection

doi: 10.3389/fcimb.2023.1268016

Figure Lengend Snippet: Intracellular generation of free radicals in CAEC exposed to culture medium (Medium), culture medium supplemented with 20% control serum (Control), 20% Post-COVID-19-serum (Post-COVID), 20% Post-COVID-19 serum supplemented with N-Acetylcysteine 1 mmol/L (Post-COVID+NAC), or Post-COVID-19 serum with Sulodexide 0.5 LRU/mL (Post-COVID+Sul).

Article Snippet: During the experiments, the primary cultures of human coronary artery endothelial cells (CAEC) obtained from Cell Applications, Inc. (San Diego, California, USA) were used.

Techniques: Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Protective role of N-acetylcysteine and Sulodexide on endothelial cells exposed on patients’ serum after SARS-CoV-2 infection

doi: 10.3389/fcimb.2023.1268016

Figure Lengend Snippet: Synthesis of IL-6 (A) and vWF (B) in CAEC exposed to culture medium (Medium), culture medium supplemented with 20% control serum (Control), 20% Post-COVID-19 serum (Post-COVID), 20% Post-COVID-19 serum supplemented with N-Acetylcysteine 1 mmol/L (Post-COVID+NAC), or Post-COVID-19 serum with Sulodexide 0.5 LRU/mL (Post-COVID+Sul).

Article Snippet: During the experiments, the primary cultures of human coronary artery endothelial cells (CAEC) obtained from Cell Applications, Inc. (San Diego, California, USA) were used.

Techniques: Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Protective role of N-acetylcysteine and Sulodexide on endothelial cells exposed on patients’ serum after SARS-CoV-2 infection

doi: 10.3389/fcimb.2023.1268016

Figure Lengend Snippet: Synthesis of tPA (A) and PAI-1 (B) in CAEC exposed to culture medium (Medium), culture medium supplemented with 20% control serum (Control), 20% Post-COVID-19 serum (Post-COVID), 20% Post-COVID-19 serum supplemented with N-Acetylcysteine 1 mmol/L (Post-COVID+NAC),or Post-COVID-19 serum with Sulodexide 0.5 LRU/mL (Post-COVID+Sul).

Article Snippet: During the experiments, the primary cultures of human coronary artery endothelial cells (CAEC) obtained from Cell Applications, Inc. (San Diego, California, USA) were used.

Techniques: Control

Journal: Nature Communications

Article Title: Endothelial Gata5 transcription factor regulates blood pressure

doi: 10.1038/ncomms9835

Figure Lengend Snippet: ( a ) GATA5 is expressed in human cardiac microvascular (CM), coronary artery (CA), dermal microvascular (DM) and pulmonary microvascular (PM) endothelial cells. ( b , c ) The vasoconstrictor response of Gata5 -null mice mesenteric arteries to norepinephrine is unaltered ( n =7 per group), while the vasodilatory response to acetylcholine is decreased ( n =10–11 per group). The results are reported as mean±s.e.m. * P <0.05 versus controls (comparison of best-fit values—effector concentration for half-maximum response (EC 50 ) and Hill slope—using an F-test). ( d – f ) Deletion of Gata5 in endothelial (e Gata5 -null mice; n =4–5 per group) but not smooth muscle cells (sm Gata5 -null mice; n =5 per group) decreases mesenteric arteries sensitivity to acetylcholine and increases BP (e Gata5 -null mice n =6–10 per group; sm Gata5 -null mice n =6–9 per group). The results are reported as mean±s.e.m. * P <0.05 versus controls (two-factor ANOVA). ( g ) The vasodilatory response of Gata5 -null mice to diethylamine NONOate, an NO donor, is unaltered ( n =7 per group). The results are reported as mean±s.e.m. (comparison of best-fit values—EC 50 and Hill slope—using an F-test). ( h , i ) NOS3 and Akt phosphorylation are decreased in Gata5 -null mice mesenteric arteries. PTEN and PDK1 phosphorylation and expression are unaltered ( n =5–7 per group). Phosphorylated proteins are normalized to total proteins. Total proteins are normalized to actin. The results are reported as mean±s.e.m. * P <0.05 versus Gata5 +/+ mice ( t -test). ( j ) Quantification of protein nitrotyrosination in mesenteric arteries of Gata5 -null mice and their controls as measured by ELISA. 3-Nitrotyrosine content is expressed as picomole of nitrotyrosine per milligram of protein ( n =5–7 per group). The results are reported as mean±s.e.m. ( t -test).

Article Snippet: Human primary endothelial cells were purchased from Promocell: Human Coronary artery endothelial cells (ref C-14022, Lot: 91001010.7P); Human Cardiac Microvascular Endothelial Cells (ref C-14029, Lot: 20224401P); Human Pulmonary Microvascular Endothelial Cells (ref C-14027, Lot: 9090301P) and HDMECs (ref C-14016, Lot: 4060603P).

Techniques: Concentration Assay, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Endothelial Gata5 transcription factor regulates blood pressure

doi: 10.1038/ncomms9835

Figure Lengend Snippet: ( a ) GATA5 expression is significantly decreased in human dermal microvascular endothelial cells infected with a lentiviral vector containing an anti- GATA5 shRNA (HDMEC-GATA5-KD). Control cells were infected with a vector containing a control shRNA (targets no known mammalian gene) (HDMEC-pLKO-Ctrl, referred here as Ctrl). ( b ) Heatmap representation of the differentially regulated genes between HDMEC-GATA5-KD cells and their controls as identified by transcriptomic analysis. Colour is function of Log2 RMA (Affymetrix microarray, n =3 per group). ( c ) Functional analysis of the differentially regulated genes between HDMEC-GATA5-KD cells and their controls. Protein kinase A pathway is the most significantly enriched pathway. Fisher's exact test P value. ( d ) Validation by qPCR (upper panel) of genes predicted by microarray (lower panel) to be up- and downregulated in HDMEC-GATA5-KD endothelial cells. ( n =5 wells per condition). Downregulated genes: PRKACB codes for the PKA catalytic subunit β, PRKAR2B for the PKA regulator subunit 2β and PRKAA2 for the AMPK catalytic subunit α2. Upregulated genes: ICAM1 codes for the intercellular adhesion molecule 1, BMP4 for the bone morphogenetic protein 4 and IL6 for the interleukin 6. The results are reported as mean±s.e.m. ** P <0.01 versus Ctrl ( t -test). ( e ) Western blot representation of phospho-NOS3, NOS3 (Ser1177) and phospho-(Ser/Thr) PKA substrate motif in HDMEC-GATA5-KD cells and their controls. ( f ) Phosphorylation of NOS3 on Ser1177 is decreased in HDMEC-GATA5-KD cells (performed twice, 2–3 wells per condition). Phospho-NOS3 is normalized to total NOS3. NOS3 is normalized to actin. The results are reported as mean±s.e.m. * P <0.05 versus Ctrl (Mann–Whitney test). ( g ) Phosphorylation of (Ser/Thr) PKA substrate motif is decreased in HDMEC-GATA5-KD cells (performed twice, 2–3 wells per condition). Phospho-(Ser/Thr) PKA substrate motif (between 25 and 250 kDa) is normalized to actin. The results are reported as mean±s.e.m. * P <0.05 versus Ctrl (Mann–Whitney test). ( h ) Western blot representation of phospho-(Ser/Thr) PKA substrate motif in mesenteric arteries of Gata5 -null mice and their controls. ( i ) In mesenteric arteries of Gata5 -null mice, there is a trend to decrease in the (Ser/Thr) PKA substrate motif phosphorylation ( n =4–5 per group). Phospho-(Ser/Thr) PKA substrate motif (between 25 and 250 kDa) is normalized to actin. The results are reported as mean±s.e.m. (Mann–Whitney test).

Article Snippet: Human primary endothelial cells were purchased from Promocell: Human Coronary artery endothelial cells (ref C-14022, Lot: 91001010.7P); Human Cardiac Microvascular Endothelial Cells (ref C-14029, Lot: 20224401P); Human Pulmonary Microvascular Endothelial Cells (ref C-14027, Lot: 9090301P) and HDMECs (ref C-14016, Lot: 4060603P).

Techniques: Expressing, Infection, Plasmid Preparation, shRNA, Microarray, Functional Assay, Western Blot, MANN-WHITNEY