и внутренней грудной  (Cell Applications Inc)

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Cytokine profiling of the enriched (6-fold), serum-free cell culture medium withdrawn from HCAECs ( top ) and HITAECs ( bottom ) treated with control DPBS, magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. Specific dot blotting kits for the measurement of cytokines. Below are the following colours that demarcate the signal from the respective antibodies which indicate overexpressed cytokines in the respective experimental groups: dark green: serpin E1/plasminogen activator inhibitor 1 (PAI-1); blue: chemokine (C-X-C motif) ligand 1/growth regulated protein alpha (CXCL1/GROα); gray: CD105/endoglin; red: interleukin-6 (IL-6); apple green: soluble interleukin 1 receptor-like 1/suppression of tumorigenicity 2 (ST2); brown: platelet-derived growth factor AB/BB (PDGF-AB/BB); gold: pentraxin-3; light blue: vascular cell adhesion molecule 1 (VCAM-1)/cluster of differentiation 106 (CD106); aquamarine: chemokine (C-C motif) ligand 5 (CCL5)/regulated on activation, normal T cell expressed and secreted (RANTES); violet: thrombospondin-1; dark pink: urokinase plasminogen activator surface receptor (uPAR); dark violet: macrophage inflammatory protein-3 alpha/CCL20; lime green: epidermal growth factor (EGF); light brown: CD147/extracellular matrix metalloproteinase inducer (EMMPRIN)/basigin; light green: CXCL5/epithelial neutrophil-activating protein 78 (ENA-78); lavender blue: hepatocyte growth factor (HGF); light violet: monocyte chemoattractant protein 1/chemokine (C-C motif) ligand 2 (MCP-1/CCL2); peach: angiogenin; orange: MCP-3/CCL7; salad green: transferrin receptor protein 1 (TfR1)/CD71; lavender: angiopoietin-2; bright green: macrophage colony-stimulating factor (M-CSF/CSF-1); forest green: macrophage migration inhibitory factor (MIF); pink: fibroblast growth factor 19 (FGF-19); baby blue: cystatin C; purple: interferon gamma-induced protein 10 (IP-10)/CXCL10; turquoise: MIP-3β; light pink: granulocyte-macrophage colony-stimulating factor (GM-CSF/CSF-2); deep pink: matrix metalloproteinase 9 (MMP-9). HCAEC: short, medium, and long arrows indicate fold change from 1.20 to 1.34, from 1.35 to 1.49, and ≥1.50, respectively, as compared with the DPBS group. HITAEC: short, medium, and long arrows indicate fold change from 1.25 to 1.34, from 1.35 to 1.49, and ≥1.50, respectively, as compared with the DPBS group.

Article Snippet: For the secretome profiling and enzyme-linked immunosorbent assay (ELISA), confluent (≈0.5 × 10 6 cells per well of 6-well plate) cultures of HCAECs (300K-05a, Cell Applications, San Diego, CA, USA) and HITAECs (308K-05a, Cell Applications, San Diego, CA, USA) in a serum-free EndoLife Medium (EL1, AppScience Products, Moscow, Russia) were incubated with 100 μL of MPPs, CPP-P, or CPP-S (0.6 × 10 5 particles per mL or 25 μg/μL calcium) or an equal volume of DPBS without Ca 2+ and Mg 2+ ions (pH = 7.4, 1.2.4.7, BioLot, St. Petersburg, Russia) for 24 h ( n = 3 wells per group).

Techniques: Cell Culture, Control, Derivative Assay, Activation Assay, Migration

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Expression and pathway enrichment analysis of protein abundance in cell culture supernatants collected from primary human coronary artery endothelial cells (HCAEC) and ( B ) primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Heat map showing the protein abundance of the 50 most abundant proteins identified in the physiological secretome (i.e., among those profiled in the cell culture supernatant from DPBS- and MPP-treated HCAECs and HITAECs, left side) and the pathological secretome (i.e., among those profiled in the cell culture supernatant from CPP-P- and CPP-S-treated HCAECs and HITAECs, right side). Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”). Orange dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in HCAECs; red dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in HITAECs; yellow dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in HCAECs; green dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in HITAECs; violet dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in both HCAECs and HITAECs; blue dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in both HCAECs and HITAECs; ( B ) Differentially expressed molecular terms were identified among these proteins during the screening of Gene Ontology and Reactome resources through the Database for Annotation, Visualization, and Integrated Discovery (DAVID).

Article Snippet: For the secretome profiling and enzyme-linked immunosorbent assay (ELISA), confluent (≈0.5 × 10 6 cells per well of 6-well plate) cultures of HCAECs (300K-05a, Cell Applications, San Diego, CA, USA) and HITAECs (308K-05a, Cell Applications, San Diego, CA, USA) in a serum-free EndoLife Medium (EL1, AppScience Products, Moscow, Russia) were incubated with 100 μL of MPPs, CPP-P, or CPP-S (0.6 × 10 5 particles per mL or 25 μg/μL calcium) or an equal volume of DPBS without Ca 2+ and Mg 2+ ions (pH = 7.4, 1.2.4.7, BioLot, St. Petersburg, Russia) for 24 h ( n = 3 wells per group).

Techniques: Expressing, Quantitative Proteomics, Cell Culture, Saline, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Pathway enrichment analysis of secretomes from primary human coronary artery endothelial cells (HCAEC) and primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Differentially expressed molecular terms identified among the proteins which are downregulated upon the incubation with CPP-P or CPP-S during the screening of Gene Ontology and Reactome resources through the Database for Annotation, Visualization, and Integrated Discovery (DAVID); ( B ) Heat map showing the downregulated expression of the ECM components (in particular basement membrane proteins) and notable and stable upregulation of soluble CD59 in HCAECs and HITAECs exposed to CPP-P or CPP-S. Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”).

Article Snippet: For the secretome profiling and enzyme-linked immunosorbent assay (ELISA), confluent (≈0.5 × 10 6 cells per well of 6-well plate) cultures of HCAECs (300K-05a, Cell Applications, San Diego, CA, USA) and HITAECs (308K-05a, Cell Applications, San Diego, CA, USA) in a serum-free EndoLife Medium (EL1, AppScience Products, Moscow, Russia) were incubated with 100 μL of MPPs, CPP-P, or CPP-S (0.6 × 10 5 particles per mL or 25 μg/μL calcium) or an equal volume of DPBS without Ca 2+ and Mg 2+ ions (pH = 7.4, 1.2.4.7, BioLot, St. Petersburg, Russia) for 24 h ( n = 3 wells per group).

Techniques: Saline, Incubation, Expressing, Membrane, Quantitative Proteomics, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Heterogeneity of protein profiles in cell culture supernatants collected from primary human coronary artery endothelial cells (HCAEC) and primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Principal component analysis performed across all experimental groups; ( B ) Principal component analysis of pooled DPBS- and MPP-treated HCAECs and HITAECs; ( C ) Venn diagrams showing the number of overlapping and unique differentially expressed proteins between HCAECs vs. HITAECs according to the various intergroup comparisons; ( D ) Volcano plot showing upregulated and downregulated proteins between pooled DPBS- and MPP-treated HCAECs and HITAECs; the dashed lines indicate logarithmic fold change and p value thresholds; selected proteins are presented below on the heat map; ( E ) Heat map demonstrating protein abundance across the 10 most upregulated and the 10 most downregulated proteins between pooled DPBS- and MPP-treated HCAECs and HITAECs. Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”).

Article Snippet: For the secretome profiling and enzyme-linked immunosorbent assay (ELISA), confluent (≈0.5 × 10 6 cells per well of 6-well plate) cultures of HCAECs (300K-05a, Cell Applications, San Diego, CA, USA) and HITAECs (308K-05a, Cell Applications, San Diego, CA, USA) in a serum-free EndoLife Medium (EL1, AppScience Products, Moscow, Russia) were incubated with 100 μL of MPPs, CPP-P, or CPP-S (0.6 × 10 5 particles per mL or 25 μg/μL calcium) or an equal volume of DPBS without Ca 2+ and Mg 2+ ions (pH = 7.4, 1.2.4.7, BioLot, St. Petersburg, Russia) for 24 h ( n = 3 wells per group).

Techniques: Cell Culture, Saline, Quantitative Proteomics, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Secretome-wide comparison of protein profiles in cell culture supernatants collected from primary human coronary artery endothelial cells (HCAEC, top) and primary human internal thoracic artery endothelial cells (HITAEC, bottom) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Principal component analysis showing the relative distance between the groups; ( B ) Venn diagrams demonstrating the numbers of differentially expressed proteins between the comparisons.

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Comparison, Cell Culture, Saline

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Number of overexpressed and underexpressed proteins in the secretomes of primary human coronary artery endothelial cells (HCAEC, top) and primary human internal thoracic artery endothelial cells (HITAEC, bottom) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), and secondary calciprotein particles (CPP-S) for 24 h. Differentially expressed proteins were defined as those with logarithmic fold change ≥ 1 and false discovery rate-corrected p value ≤ 0.05. All comparisons (CPP-P vs. DPBS, CPP-P vs. MPP, CPP-S vs. DPBS, CPP-S vs. MPP, MPP vs. DPBS, and CPP-S vs. CPP-P) for each cell line were analysed.

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Saline

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Expression and pathway enrichment analysis of protein abundance in cell culture supernatants collected from primary human coronary artery endothelial cells (HCAEC) and ( B ) primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Heat map showing the protein abundance of the 50 most abundant proteins identified in the physiological secretome (i.e., among those profiled in the cell culture supernatant from DPBS- and MPP-treated HCAECs and HITAECs, left side) and the pathological secretome (i.e., among those profiled in the cell culture supernatant from CPP-P- and CPP-S-treated HCAECs and HITAECs, right side). Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”). Orange dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in HCAECs; red dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in HITAECs; yellow dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in HCAECs; green dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in HITAECs; violet dots highlight the proteins which are exclusively presented among the 50 most abundant in the physiological secretome in both HCAECs and HITAECs; blue dots highlight the proteins which are exclusively presented among the 50 most abundant in the pathological secretome in both HCAECs and HITAECs; ( B ) Differentially expressed molecular terms were identified among these proteins during the screening of Gene Ontology and Reactome resources through the Database for Annotation, Visualization, and Integrated Discovery (DAVID).

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Expressing, Quantitative Proteomics, Cell Culture, Saline, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Pathway enrichment analysis of secretomes from primary human coronary artery endothelial cells (HCAEC) and primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Differentially expressed molecular terms identified among the proteins which are downregulated upon the incubation with CPP-P or CPP-S during the screening of Gene Ontology and Reactome resources through the Database for Annotation, Visualization, and Integrated Discovery (DAVID); ( B ) Heat map showing the downregulated expression of the ECM components (in particular basement membrane proteins) and notable and stable upregulation of soluble CD59 in HCAECs and HITAECs exposed to CPP-P or CPP-S. Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”).

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Saline, Incubation, Expressing, Membrane, Quantitative Proteomics, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: The 10 most downregulated and 10 most upregulated proteins in secretomes from primary human coronary artery endothelial cells (HCAEC) and primary human internal thoracic artery endothelial cells (HITAEC) treated with primary calciprotein particles (CPP-P) or secondary calciprotein particles (CPP-S) in comparison with those incubated with Dulbecco’s phosphate-buffered saline (DPBS) or magnesiprotein particles (MPP), for 24 h. N/A means non-inclusion in the top 10 most downregulated or upregulated proteins, but not the absence of differential expression. Differentially expressed proteins were defined as those with logarithmic fold change ≥ 1 and false discovery rate-corrected p value ≤ 0.05. Log 2 fold change means logarithmic fold change.

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Comparison, Incubation, Saline, Quantitative Proteomics

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Analysis of protein–protein interactions among the downregulated proteins in secretomes from primary human coronary artery endothelial cells (HCAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) CPP-P vs. DPBS; ( B ) CPP-P vs. MPP; ( C ) CPP-S vs. DPBS; ( D ) CPP-S vs. MPP. Analysis was performed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING).

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Protein-Protein interactions, Saline

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Analysis of protein–protein interactions among the downregulated proteins in secretomes from primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) CPP-P vs. DPBS; ( B ) CPP-P vs. MPP; ( C ) CPP-S vs. DPBS; ( D ) CPP-S vs. MPP. Analysis was performed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING).

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Protein-Protein interactions, Saline

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Heterogeneity of protein profiles in cell culture supernatants collected from primary human coronary artery endothelial cells (HCAEC) and primary human internal thoracic artery endothelial cells (HITAEC) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. ( A ) Principal component analysis performed across all experimental groups; ( B ) Principal component analysis of pooled DPBS- and MPP-treated HCAECs and HITAECs; ( C ) Venn diagrams showing the number of overlapping and unique differentially expressed proteins between HCAECs vs. HITAECs according to the various intergroup comparisons; ( D ) Volcano plot showing upregulated and downregulated proteins between pooled DPBS- and MPP-treated HCAECs and HITAECs; the dashed lines indicate logarithmic fold change and p value thresholds; selected proteins are presented below on the heat map; ( E ) Heat map demonstrating protein abundance across the 10 most upregulated and the 10 most downregulated proteins between pooled DPBS- and MPP-treated HCAECs and HITAECs. Protein abundance is represented as log 2 -transformed, imputed, and normalised data (see . in Materials and Methods “Proteomic profiling”).

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Cell Culture, Saline, Quantitative Proteomics, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Endothelial Cell Secretomes After Exposure to Calciprotein Particles Reveals Downregulation of Basement Membrane Assembly and Increased Release of Soluble CD59

doi: 10.3390/ijms252111382

Figure Lengend Snippet: Enzyme-linked immunosorbent assay (ELISA) measurements of the levels of protectin (CD59), osteonectin (secreted protein acidic and rich in cysteine, SPARC), perlecan (heparan sulfate proteoglycan 2, HSPG2), and fibronectin (FN1) in the pre-centrifuged, serum-free cell culture supernatant (2000× g ) from primary human coronary artery endothelial cells (HCAEC, top ) and primary human internal thoracic artery endothelial cells (HITAEC, bottom ) treated with Dulbecco’s phosphate-buffered saline (DPBS), magnesiprotein particles (MPP), primary calciprotein particles (CPP-P), or secondary calciprotein particles (CPP-S) for 24 h. Blue, violet, pink and red dots are for the DPBS, MPP, CPP-P, and CPP-S-treated cells, respectively. Each dot on the plots represents one measurement ( n = 12 measurements per group). Whiskers indicate the range, box bounds indicate the 25th–75th percentiles, and centre lines indicate the median. p values are provided above boxes, Kruskal–Wallis test with Dunn’s multiple comparisons test.

Article Snippet: Primary cultures of human coronary artery endothelial cells (HCAECs, 300K-05a, Cell Applications, San Diego, CA, USA) and human internal thoracic artery endothelial cells (HITAECs, 308K-05a, Cell Applications, San Diego, CA, USA) were grown in T-75 flasks (N-708003, Wuxi NEST Biotechnology Co., Ltd., Wuxi, China) according to the manufacturer’s protocols using EndoBoost Medium (EB1, AppScience Products, Moscow, Russia) until reaching confluence.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Saline