Journal: Cells

Article Title: Sphingosylphosphorylcholine (SPC), a Causative Factor of SPC-Induced Vascular Smooth Muscle Cells Contraction, Is Taken Up via Endocytosis

doi: 10.3390/cells12020265

Figure Lengend Snippet: Analysis of sphingosylphosphorylcholine (SPC) and microdomain involvement in SPC-induced contractions of human coronary artery smooth muscle cells (HCASMCs). ( a ) Expression levels of the microdomain markers flotillin-1 (Flot1) and caveolin-1 (Cav1). Black and white indicate Cav1 and Flot1, respectively. ( b ) HCASMCs treated with 5 mM (top row) or 10 mM (bottom row) of methyl-β-cyclodextrin (MβCD). The left and right columns depict unstimulated and SPC-stimulated cells, respectively. ( c ) Contracted cell rates with microdomains disrupted by 5 mM of MβCD. * p < 0.05 vs. without SPC stimulation and fisetin treatment or only SPC stimulation. Data are represented as mean ± standard deviation.

Article Snippet: Normal HCASMCs (Kurabo, Osaka, Japan) recovered from a male were cultured in HuMedia-SG2 growth medium (Kurabo, Osaka, Japan) in a humidified atmosphere at 37 °C containing 5% CO 2 (PHC, Tokyo, Japan) until 80–90% confluency.

Techniques: Expressing, Standard Deviation

Journal: Cells

Article Title: Sphingosylphosphorylcholine (SPC), a Causative Factor of SPC-Induced Vascular Smooth Muscle Cells Contraction, Is Taken Up via Endocytosis

doi: 10.3390/cells12020265

Figure Lengend Snippet: Localization of sphingosylphosphorylcholine (SPC) in human coronary artery smooth muscle cells (HCASMCs). ( a ) Representative image of NBD-SPC (green) localization in abnormally contracting HCASMCs. DAPI (blue), nuclear stain; PlasMem Bright Red (Red), plasma membrane stain. The top and bottom rows depict untreated control (Ctrl) cells and fisetin-treated cells. ( b ) Percentage of positive cells for NBD-SPC. Black and white indicate the number of abnormally contracting and non-contracting cells. ( c ) Mean fluorescence intensity of NBD-SPC. ( d ) Flow cytometric analysis of NBD-SPC-positive cells treated with 5 mM of MβCD. * p < 0.05 vs. 0 μM NBD-SPC-treated cells and a: p < 0.05 vs. abnormally contracting cells. N.D. = not detected. Data are represented as mean ± standard deviation.

Article Snippet: Normal HCASMCs (Kurabo, Osaka, Japan) recovered from a male were cultured in HuMedia-SG2 growth medium (Kurabo, Osaka, Japan) in a humidified atmosphere at 37 °C containing 5% CO 2 (PHC, Tokyo, Japan) until 80–90% confluency.

Techniques: Staining, Clinical Proteomics, Membrane, Control, Fluorescence, Standard Deviation

Journal: Cells

Article Title: Sphingosylphosphorylcholine (SPC), a Causative Factor of SPC-Induced Vascular Smooth Muscle Cells Contraction, Is Taken Up via Endocytosis

doi: 10.3390/cells12020265

Figure Lengend Snippet: The number of exosomes in human coronary artery smooth muscle cells (HCASMCs). ( a ) (1) Flow cytometric dot plots representing CD9- and CD63-positive populations. (2) Number of exosomes generated from control (Ctrl) cells and fisetin-treated cells. ( b ) Number of exosomes in HCASMCs relative to unstimulated Ctrl cells. * p < 0.05 vs. Ctrl without stimulation. # p < 0.05 vs. without sphingosylphosphorylcholine (SPC) stimulation at the same time. Data are expressed as mean ± standard deviation.

Article Snippet: Normal HCASMCs (Kurabo, Osaka, Japan) recovered from a male were cultured in HuMedia-SG2 growth medium (Kurabo, Osaka, Japan) in a humidified atmosphere at 37 °C containing 5% CO 2 (PHC, Tokyo, Japan) until 80–90% confluency.

Techniques: Generated, Control, Standard Deviation

Journal: Biotechnology Research International

Article Title: Human Coronary Artery Smooth Muscle Cell Responses to Bioactive Polyelectrolyte Multilayer Interfaces

doi: 10.4061/2011/854068

Figure Lengend Snippet: Morphology of hCASMC cultures on PEMU surfaces after 24 hours on (a) uncoated glass, (b) (PFPVP/Nafion) 2 , (c) (PDADMA/PSS) 2 , and (d) (PAH/PAA- co -PAEDAPS) 2 . After 5 days, hCASMCs on (e) uncoated glass achieved a uniform, confluent layer, while cells on (f) (PFPVP/Nafion) 2 exhibited a “hill and valley” morphology. Cells on (g) (PDADMA/PSS) 2 exhibited continued aggregation with a pattern similar to that seen on (PFPVP/Nafion) 2 , but with less alignment of the cells. Cultures on (h) (PAH/PAA- co -PAEDAPS) 2 were virtually unchanged after 5 days. All images are 100x. Scale bar = 20 μ m.

Article Snippet: Human coronary artery smooth muscle cells (hCASMCs) (BioWhittaker, Inc.) were routinely cultured in Smooth Muscle Basal Medium (SmBM, Cambrex) in which 475 mL of SmBM was supplemented with 0.5 mL human epidermal growth factor (hEGF), 0.5 mL insulin, 1.0 mL human fibroblast growth factor B (hFGF-B) (all from Cambrex at 1 μ g/mL, SmGM-2 SingleQuots), 25 mL (5%) fetal bovine serum (FBS) (Cambrex), and 0.5 mL gentamicin/amphotericin-B (GA-1000) (Cambrex).

Techniques:

Journal: International Journal of General Medicine

Article Title: Phosphoglycerate Dehydrogenase Overexpression Inhibits Ferroptosis to Repress Calcification of Human Coronary Artery Vascular Smooth Muscle Cells via the P53/SLC7A11 Pathway

doi: 10.2147/IJGM.S473908

Figure Lengend Snippet: PHGDH overexpression inhibited the calcification level of calcifying medium (CM)-induced human coronary artery smooth muscle cells (HCASMCs). ( A ) T he relative mRNA and protein expression levels of PHGDH. ( B ) The cell viability. ( C ) The results of alizarin red staining. ( D ) The protein expression level of RUNX2 and bone morphogenetic protein 2 (BMP2). ( E ) The relative ALP activity. ( F ) The calcium content. **p < 0.01 vs the Normal group; ## p < 0.01 vs the oe-NC group.

Article Snippet: Primary HCASMCs were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Over Expression, Expressing, Staining, Activity Assay

Journal: International Journal of General Medicine

Article Title: Phosphoglycerate Dehydrogenase Overexpression Inhibits Ferroptosis to Repress Calcification of Human Coronary Artery Vascular Smooth Muscle Cells via the P53/SLC7A11 Pathway

doi: 10.2147/IJGM.S473908

Figure Lengend Snippet: Inhibition of ferroptosis suppressed the calcification level of CM-induced HCASMCs. ( A ) The results of alizarin red staining. Fer-1: ferrostatin-1. ( B ) The calcium content. ( C ) The protein expression level of RUNX2 and BMP2. ( D ) The expression level of glutathione (GSH). ( E ) The protein expression levels of cyclooxygenase 2 (COX2) and glutathione peroxidase 4 (GPX4). **p < 0.01 vs the Normal group; ## p < 0.01 vs the Control group.

Article Snippet: Primary HCASMCs were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Inhibition, Staining, Expressing, Control

Journal: International Journal of General Medicine

Article Title: Phosphoglycerate Dehydrogenase Overexpression Inhibits Ferroptosis to Repress Calcification of Human Coronary Artery Vascular Smooth Muscle Cells via the P53/SLC7A11 Pathway

doi: 10.2147/IJGM.S473908

Figure Lengend Snippet: PHGDH overexpression inhibited the calcification level of CM-induced HCASMCs by inhibiting ferroptosis. ( A ) The results of alizarin red staining. ( B ) The calcium content. ( C ) The relative ALP activity. ( D ) The expression level of GSH. (E) The protein expression level of COX2 and GPX4. **p < 0.01.

Article Snippet: Primary HCASMCs were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Over Expression, Staining, Activity Assay, Expressing