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: 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

Journal:

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

doi: 10.2353/ajpath.2006.050284

Figure Lengend Snippet: A and B: Expression of BMP-2 in endothelial and smooth muscle cells microdissected from coronary arteries (A) and in cultured primary rat CAECs and VSMCs (B). Analysis of mRNA expression was performed by real-time QRT-PCR. GAPDH was used for normalization. Data are mean ± SEM (n = 3 to 5 for each group). *P < 0.05. C: Fluorescent photomicrographs showing that immunolabeling for BMP-2 (green) is present in endothelial cells (double arrows) and medial smooth muscle cells (bold arrow, smooth muscle cells show red immunofluorescent labeling for α-smooth muscle actin). D: VISTA plot showing the percentage of conservation between the mRNAs transcribed from the rat BMP-2 and BMP-4 genes. E and F: Expression of BMP-4 in endothelial and smooth muscle cells microdissected from coronary arteries (E) and in cultured CAECs and VSMCs (F). Data are mean ± SEM (n = 3 to 5 for each group). *P < 0.05.

Article Snippet: 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: Expressing, Cell Culture, Quantitative RT-PCR, Immunolabeling, Labeling

Journal:

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

doi: 10.2353/ajpath.2006.050284

Figure Lengend Snippet: Effect of TNF-α (1 or 10 ng/ml, for 24 hours) on the expression of BMP-2 mRNA (A, C: QRT-PCR) and protein (B, D: Western blotting) in cultured primary rat CAECs and VSMCs. Data are mean ± SEM (n = 4 to 8 for each group). *P < 0.05 versus untreated control. E: Expression of BMP-2 mRNA in endothelial and smooth muscle cells microdissected from control and TNF-α-treated (10 ng/ml, for 24 hours) cultured rat aortas. Data are mean ± SEM. *P < 0.05 versus untreated control. F: Reporter gene assay showing the effects of TNF-α (10 ng/ml) on NF-κΒ reporter activity in CAECs and VSMCs. 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 seven independent transfections. In control experiments PDTC was used to inhibit NF-κB activity. Data are mean ± SEM. *P < 0.05 versus control. G: Effect of TNF-α (1 or 10 ng/ml, for 24 hours) on the expression of BMP-4 mRNA (QRT-PCR) in CAECs and VSMCs (H). Data are mean ± SEM. *P < 0.05 versus untreated control.

Article Snippet: 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: Expressing, Quantitative RT-PCR, Western Blot, Cell Culture, Reporter Gene Assay, Activity Assay, Transfection, Luciferase, Construct

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: 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