Journal: International Journal of Molecular Sciences

Article Title: High-Content Imaging and Machine Learning Classify Phenotypical Change in Coronary Artery Endothelial Cells Caused by BPS

doi: 10.3390/ijms27073259

Figure Lengend Snippet: Representative high-content microscopy images of human coronary artery endothelial cells (HCAEC) exposed to vehicle control (CTRL) or 0.1 µM Bisphenol S (BPS) for 96 h and stained using the PhenoVue Cell Painting assay. For each condition, a representative field acquired at 40× magnification and a higher-magnification inset are shown. Rows correspond to the individual fluorescence channels: Hoechst 33342 (nuclei), PhenoVue Fluor 488 Concanavalin A (endoplasmic reticulum and intracellular membranes), PhenoVue 512 nucleic acid stain (RNA/nucleoli), PhenoVue Fluor 555 wheat germ agglutinin (plasma membrane), PhenoVue 641 mitochondrial stain (mitochondria), and the merged image. White boxes represent the part of the image used for the related inset. Scale bar: 50 µm, 40× objective.

Article Snippet: Primary human coronary artery endothelial cells (HCAEC; ATCC ® PCS-100-020TM, Innovation, VA, USA) were cultured according to the supplier’s recommendations.

Techniques: Microscopy, Control, Staining, Fluorescence, Clinical Proteomics, Membrane

Journal: Advanced Healthcare Materials

Article Title: Nitric Oxide‐Releasing Catheters with Phenol‐Amine Catalytic Coatings for Improved Anti‐Inflammatory Performance

doi: 10.1002/adhm.202500457

Figure Lengend Snippet: a) HCASMCs viability, b) number of cells, and c) endogenous NO generation after incubation with uncoated and coated catheter segments compared to the blank group, measured using the Live/Dead assay, Hoechst staining, and DAF‐FM diacetate, respectively, at i) 48 h and ii) 72 h. Statistical significance relative to control tests was calculated using one‐way ANOVA, ns = not significant, * p < 0.1, ** p < 0.01, **** p < 0.0001. n = 6; error bars represent standard deviation.

Article Snippet: Human coronary artery smooth muscle cells (HCASMCs) and smooth muscle cell growth medium kit were purchased from Cell Applications.

Techniques: Incubation, Live Dead Assay, Staining, Control, Standard Deviation

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Glutamate Exacerbates Traumatic Brain Injury‐Induced Acute Lung Injury Through NMDAR / ROS /Ca 2+ Signaling Pathway in Pulmonary Endothelial Cells

doi: 10.1002/kjm2.70087

Figure Lengend Snippet: Changes of serious lung injury after TBI‐ALI. (A) The TTC staining was performed to evaluate the brain injury volume. (B) The level of cytokines (TNF‐α, IL‐1β, and IL‐6) in peripheral blood. (C) The level of cytokines (TNF‐α, IL‐1β, and IL‐6) in BALF. (D, E) The wet/dry weight ratio and the protein concentration are detected. (F, G) Corresponding lung H&E staining and acute lung injury scores. (H) The expression of GluN1 in HPMVECs. Scale bar, 200 μm. Results represent the mean ± SEM of independent experiments of animals ( n = 8). * p < 0.05 versus Sham group; # p < 0.05 versus TBI group.

Article Snippet: Human pulmonary microvascular endothelial cells (HPMVECs) were purchased from Cell Applications Inc. (San Diego, CA, USA; Catalog No. 300K‐05a) and cultured in RPMI‐1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) with 1% penicillin–streptomycin and 10% fetal bovine serum (FBS, Gbico, Grand Island, NY, USA) at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Staining, Protein Concentration, Expressing

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Glutamate Exacerbates Traumatic Brain Injury‐Induced Acute Lung Injury Through NMDAR / ROS /Ca 2+ Signaling Pathway in Pulmonary Endothelial Cells

doi: 10.1002/kjm2.70087

Figure Lengend Snippet: Glutamate alters NMDAR/ROS/Ca 2+ pathway. (A) Cell viability (percentage of untreated control) of HPMVECs after the treatment of glutamate. (B) Immunofluorescence images showing ROS production in HPMVECs. (C) Comparison of Ca 2+ concentration in each group. (D) The levels of p‐NFAT and p‐p65 in cytoplasm and nucleus were tested by western blot. (E) Immunofluorescence stain of p‐NFAT and p‐p65 in nucleus. Results represent the mean ± SEM of independent experiments of cells ( n = 3). * p < 0.05 versus Sham group; # p < 0.05 versus Glu group.

Article Snippet: Human pulmonary microvascular endothelial cells (HPMVECs) were purchased from Cell Applications Inc. (San Diego, CA, USA; Catalog No. 300K‐05a) and cultured in RPMI‐1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) with 1% penicillin–streptomycin and 10% fetal bovine serum (FBS, Gbico, Grand Island, NY, USA) at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Control, Immunofluorescence, Comparison, Concentration Assay, Western Blot, Staining

Journal:

Article Title: Vasculoprotective Effects of Anti-Tumor Necrosis Factor-? Treatment in Aging

doi: 10.2353/ajpath.2007.060708

Figure Lengend Snippet: Concentration dependence of the vascular effects of TNF-α. A and B: Superoxide production (A; measured by the lucigenin chemiluminescence method) and relaxations to acetylcholine (B) and in ring preparations of carotid arteries of young F344 rats maintained in vessel culture (for 24 hours) in the absence and presence of TNF-α. Data are mean ± SEM (n = 4 to 6 in each group) *P < 0.05. C: DNA fragmentation in arteries of young F344 rats maintained in vessel culture (for 24 hours) in the absence and presence of TNF-α. Data are mean ± SEM (n = 4 to 6 in each group) *P < 0.05. D: Reporter gene assay showing the effects of TNF-α on NF-κΒ reporter activity in coronary arterial endothelial cells. Endothelial cells were transiently co-transfected with NF-κΒ-driven firefly luciferase and CMV-driven Renilla luciferase constructs followed by TNF-α stimulation. Cells were then lysed and subjected to luciferase activity assay. After normalization, relative luciferase activity was obtained from four independent transfections (data are mean ± SEM, *P < 0.05 versus control). E and F: Effect of TNF-α treatment (24 hours) on the expression of iNOS in coronary arterial endothelial cells (E) and smooth muscle cells (F). Analysis of mRNA expression was performed by real-time QRT-PCR. Data are mean of four independent experiments.

Article Snippet: Primary rat coronary arterial endothelial cells (Celprogen, San Pedro, CA) and aortic smooth muscle cells (Cell Applications Inc., San Diego, CA) were treated with recombinant TNF-α (from 0.1 to 100 ng/ml) as described.

Techniques: Concentration Assay, Reporter Gene Assay, Activity Assay, Transfection, Luciferase, Construct, Expressing, Quantitative RT-PCR

Journal: Diabetes & Vascular Disease Research

Article Title: Advanced glycation end products impair coronary artery BK channels via AMPK/Akt/FBXO32 signaling pathway

doi: 10.1177/14791641231197107

Figure Lengend Snippet: Effects of inhibition of AGEs on coronary artery tensions and BK channel densities and protein expression (a) Representative tracings for 60 mmol/L KCl and 100 nmol/L IBTX induced vascular tension alterations of coronary arterial rings from C+V, DM+V, C+A and DM+A groups. (b) Graph data showing the vascular tension alterations induced by KCl. (c) Graph data showing the vascular tension alterations (IBTX/KCl). (d and e) Whole-cell potassium currents before and after application of 100 nmol/L IBTX, and the I-V relationship of IBTX-sensitive currents of control and AGEs-cultured freshly isolated rat coronary arterial SMCs ( n = 3∼6 per group). (f) The representative tracings of baseline potassium currents and potassium currents after application of 100 nM IBTX in rat coronary arterial SMCs of the C+V, DM+V, C+A and DM+A groups, respectively ( n = 3∼5 per group). (g) Graph data showing IBTX-sensitive current densities at the testing potential of +100 mV in rat coronary arterial SMCs of the four groups. (h–j) The protein expressions of BK-α and BK-β1 in human coronary arterial SMCs in the BSA and BSA-AGEs groups ( n = 6∼9 per group). Quantitative analysis of BK-α and BK-β1 were normalized to GAPDH protein expression levels. (k-l) The mRNA expression of BK-α and BK-β1 in rat coronary arteries of the C+V, DM+V, C+A and DM+A groups. β-actin was used as an internal control to normalize differences in the amount of total RNA in each rat sample ( n = 4 per group). (m and n) The mRNA expression of BK-α and BK-β1 in human coronary arterial SMCs of the NG, HG, NG+A, HG+A groups. GAPDH was used as an internal control to normalize differences in the amount of total RNA in each cell sample ( n = 4∼5 per group). (o–q) Protein expressions of BK-α and BK-β1 in rat coronary arteries of the C+V, DM+V, C+A and DM+A groups. Quantitative analysis of BK-α and BK-β1 were normalized to GAPDH protein expression levels ( n = 5 per group). (r–t) Protein expressions of BK-α and BK-β1 in human coronary arterial SMCs of the NG, HG, NG+A, HG+A groups. Quantitative analysis of BK-α and BK-β1 were normalized to GAPDH protein expression levels ( n = 5∼9 per group). (C+V: Control + Vehicle; C+A: Control + aminoguanidine; DM+V: DM + Vehicle; DM+A: DM + aminoguanidine. NG: normal glucose; HG: high glucose; NG+A: normal glucose + aminoguanidine; HG+A: high glucose + aminoguanidine).

Article Snippet: Human coronary arterial SMCs (ATCC, #PCS-100-021) and the culture medium (ATCC, #PCS-100-042 and #PCS-100-030) were purchased from ATCC.

Techniques: Inhibition, Expressing, Control, Cell Culture, Isolation

Journal: Diabetes & Vascular Disease Research

Article Title: Advanced glycation end products impair coronary artery BK channels via AMPK/Akt/FBXO32 signaling pathway

doi: 10.1177/14791641231197107

Figure Lengend Snippet: Regulation of Akt in AGEs-mediated FBXO32-induced BK-β1 degradation (a and b) Protein expression of FBXO32 in rat coronary arteries of four groups ( n = 5 per group). (c and d) Protein expression of FBXO32 in human coronary arterial SMCs of four cell groups. Quantitative analysis of FBXO32 was normalized to GAPDH protein expression levels. (e–g) Phosphorylation levels of Akt and total Akt in rat coronary arteries of four groups ( n = 8 per group). (h–j) Phosphorylation levels of Akt and total Akt in human coronary arterial SMCs of four groups ( n = 3 per group). The phosphorylation level of Akt (k and n) and the protein expressions of FBXO32 (l and o) and BK-β1 (m and p) were measured after human coronary arterial SMCs were incubated for 96 h in DMEM containing 25.5 mmol/L glucose, or 25.5 mmol/L glucose with aminoguanidine in the absence or presence of MK2206 (0.3 μM) ( n = 5∼10 per group). MK2206 was added at the beginning and remained for 6 h (C+V: Control + Vehicle; C+A: Control + aminoguanidine; DM+V: DM + Vehicle; DM+A: DM + aminoguanidine. NG: normal glucose; HG: high glucose; NG+A: normal glucose + aminoguanidine; HG+A: high glucose + aminoguanidine.)

Article Snippet: Human coronary arterial SMCs (ATCC, #PCS-100-021) and the culture medium (ATCC, #PCS-100-042 and #PCS-100-030) were purchased from ATCC.

Techniques: Expressing, Phospho-proteomics, Incubation, Control

Journal: Diabetes & Vascular Disease Research

Article Title: Advanced glycation end products impair coronary artery BK channels via AMPK/Akt/FBXO32 signaling pathway

doi: 10.1177/14791641231197107

Figure Lengend Snippet: Regulation of AMPK in Akt-mediated FBXO32-induced BK-β1 degradation by AGEs (a–c) Protein expression of p-AMPK and AMPK in rat coronary arteries from the four groups ( n = 8 per group). (d–f) Protein expression of p-AMPK and AMPK in human coronary arterial SMCs from the four groups ( n = 9 per group). Quantitative analysis of p-AMPK and AMPK was normalized to GAPDH protein expression levels. (g) Human coronary arterial SMCs were incubated for 96 h in DMEM containing 25.5 mmol/L glucose, or 25.5 mmol/L glucose and aminoguanidine in the absence or presence of Compound C (CC, 1 μM). Subsequently, the phosphorylation level of AMPK (h and i), AKT (j and k), and the protein expressions of FBXO32 (l) and BK-β1 (m) were measured ( n = 8 and 9 per group). Quantitative analysis of FBXO32 and BK-β1 was normalized to GAPDH protein expression levels.

Article Snippet: Human coronary arterial SMCs (ATCC, #PCS-100-021) and the culture medium (ATCC, #PCS-100-042 and #PCS-100-030) were purchased from ATCC.

Techniques: Expressing, Incubation, Phospho-proteomics

Journal: ACS applied materials & interfaces

Article Title: Extracellular Vesicles Enhance the Remodeling of Cell-Free Silk Vascular Scaffolds in Rat Aortae.

doi: 10.1021/acsami.0c06609

Figure Lengend Snippet: Figure 2: A: Proliferation and B: migration of smooth muscle cells (SMCs) and endothelial cells (ECs) when exposed to extracellular vesicle (EV) based treatments. BM: Basal media; PBS: Phosphate buffered saline; EV50: 50 μl of EV isolate; EV150: 150 μl of EV isolate; SBM: Supplemented basal media. * represents p < 0.05, ** represents p < 0.005 and *** represents p < 0.0001.

Article Snippet: 22 Human coronary artery endothelial cells (ECs) were obtained from Cell Applications 23 (#300-05a), cultured in supplemented basal media (#212K-500, Cell Applications 24 Inc, SBM) and used at passage 4.

Techniques: Migration, Saline

Journal:

Article Title: Bone Morphogenetic Protein-2 Induces Proinflammatory Endothelial Phenotype

doi: 10.2353/ajpath.2006.050284

Figure Lengend Snippet: A: Representative Western blot (left) and densitometric data (right) showing the effect of increasing concentrations of anti-p42/p44 siRNAs on the expression of p42/44 MAP kinase in primary human coronary arterial endothelial cells (HCAECs). B: Effect of pretreatment with anti-p42/p44 siRNAs on BMP-2- and BMP-4- (10 ng/ml, for 2 hours) induced adhesion of fluorescently labeled PMA-stimulated monocytes to HCAECs. PD98059 (30 minutes, 10 μmol/L) was used to pharmacologically inhibit MAP kinase activity. TNF-α (10 ng/ml) was used as positive control. Data are mean ± SEM. *P < 0.05 versus control, #P < 0.05 versus BMP-2/4 treatment. C–F: Representative Western blots (C, E) and densitometric data (D, F) showing the time course of p42/44 MAP kinase phosphorylation in BMP-2 (10 ng/ml)-treated HCAECs (C, D) and rat carotid arterial segments (E, F). G: Representative Western blot (top) and densitometric data (bottom) showing BMP-2 (10 ng/ml, 10 minutes)-induced phosphorylation of p42/44 MAP kinase in HCAECs pretreated with DPI, chelerythrine, and PD98059.

Article Snippet: Studies on Endothelial Cell Cultures Primary rat coronary arterial endothelial cells (CAECs; Celprogen, San Pedro, CA), rat aortic vascular smooth muscle cells (VSMCs; Cell Applications Inc., San Diego, CA), and SV-40-immortalized rat aortic smooth muscle cells (SV40-SMC, no. CRL-2018; American Type Culture Collection, Manassas, VA) were maintained in culture as described.

Techniques: Western Blot, Expressing, Labeling, Activity Assay, Positive Control