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: 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: Cells

Article Title: BMPR2 Dosage Gates BMP9/10 Signaling Output in Pulmonary Artery Endothelium

doi: 10.3390/cells15060492

Figure Lengend Snippet: BMP9 and BMP10 selectively activate SMAD1/5/8 signaling and induce proliferation in pulmonary artery endothelial cells but not pulmonary artery smooth muscle cells. ( A ) Western blot analysis of phosphorylated SMAD1/5/8 and SMAD2/3 in PAECs treated with the indicated TGF-β superfamily ligands (0.8 nM) or untreated control (UT); β-actin serves as a loading control. ( B ) PAEC proliferation measured by BrdU incorporation following ligand treatment (0.8 nM), normalized to UT. ( C ) Western blot analysis of phosphorylated SMAD1/5/8 and SMAD2/3 in PASMCs treated with the indicated ligands (0.8 nM); β-actin serves as a loading control. ( D ) PASMC proliferation measured by BrdU incorporation following ligand treatment (0.8 nM), normalized to UT. Data are shown as mean ± SD ( n = 3 replicate wells). Statistical significance was assessed by one-way ANOVA with Dunnett’s multiple-comparisons test (each ligand vs. UT). Statistical significance was assessed by one-way ANOVA with Dunnett’s multiple-comparisons test (each ligand vs. UT). ** p < 0.01, *** p < 0.001; ns, not significant.

Article Snippet: Cell Lines and Culture: Human primary pulmonary artery endothelial cells (PAECs; ATCC PCS-100-022), pulmonary artery smooth muscle cells (PASMCs; ATCC PCS-100-023), and HEK293 cells (ATCC CRL-1573) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Western Blot, Control, BrdU Incorporation Assay

Journal: Cells

Article Title: BMPR2 Dosage Gates BMP9/10 Signaling Output in Pulmonary Artery Endothelium

doi: 10.3390/cells15060492

Figure Lengend Snippet: BMPR2 dosage-dependent model for BMP9/10 signaling output in pulmonary artery endothelial cells. Schematic illustrating how BMPR2 abundance constrains BMP9/10 (ALK1-dependent) canonical signaling output and downstream cellular programs in PAECs. ( A ) BMPR2-sufficient (~100%) state: BMP9/10 predominantly signal through ALK1–BMPR2 complexes, generating pSMAD1/5/8 output consistent with a threshold-like requirement for proliferation; bimagrumab (BiMab) produces no effect detected under BMPR2-replete conditions. ( B ) BMPR2-limiting (~50%) state: Reduced BMPR2 attenuates BMP9/10-induced canonical output and is associated with reduced proliferation and increased caspase-3/7 activity consistent with stress/injury. Under BMPR2-limiting conditions, residual canonical output becomes bimagrumab-sensitive, consistent with context-dependent contribution of Activin type II receptors (predominantly ACVR2A in PAECs; see for BMP10 affinity comparisons) to the remaining pSMAD1/5/8 signal. A putative non-canonical stress-signaling arm is shown as a proposed intermediate. Solid arrows denote observed relationships; dashed arrows and dashed-outline boxes denote proposed steps. Node shading and output gauges depict relative canonical signaling output.

Article Snippet: Cell Lines and Culture: Human primary pulmonary artery endothelial cells (PAECs; ATCC PCS-100-022), pulmonary artery smooth muscle cells (PASMCs; ATCC PCS-100-023), and HEK293 cells (ATCC CRL-1573) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay

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: 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