Journal: Cell reports

Article Title: Circulating miRNA Spaceflight Signature Reveals Targets for Countermeasure Development

doi: 10.1016/j.celrep.2020.108448

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: EGM-2 Endothelial Cell Growth Medium-2 Bullet-Kit , Lonza , Cat# CC-3162.

Techniques: Clinical Proteomics, Recombinant, Saline, Multiplex Assay, cDNA Synthesis, SYBR Green Assay, Sample Prep, Multiplexing, Software, Single-cell Analysis, Real-time Polymerase Chain Reaction, Irradiation

Journal: Circulation research

Article Title: Angiogenic Mechanisms of Human CD34 + Stem Cell Exosomes in the Repair of Ischemic Hindlimb

doi: 10.1161/CIRCRESAHA.116.310557

Figure Lengend Snippet: Uptake and transfer of CD34Exo and exosomal miRNAs by endothelial cells in vitro and in vivo. Flow cytometry analysis of CD34+ cells (A) or exosomes isolated from CD34+ cells (B), transfected as indicated; confocal image of HUVECs treated with Cy3miRNA-CD34Exo (C); flow cytometry analysis of single cell suspensions from post-ischemic hindlimb tissue injected with R-PE-CD34Exo at 2h (D); % of cells uptaking exosomes (PE) from total number of each cell types quantified from Figure 5D (E), (n=3–5, *p<0.05).

Article Snippet: Human umbilical-vein endothelial cells (HUVECs) (Lonza Inc. Allendale, NJ) were maintained in endothelial growth medium-2 (EGM ™ -2; Lonza Inc.) and starved in EBM-2 medium containing 0.25% fetal bovine serum for 24 hours before cell assays were performed.

Techniques: In Vitro, In Vivo, Flow Cytometry, Isolation, Transfection, Injection

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Ectopic Fatty Acid–Binding Protein 4 Expression in the Vascular Endothelium is Involved in Neointima Formation After Vascular Injury

doi: 10.1161/JAHA.117.006377

Figure Lengend Snippet: Overexpression and secretion of fatty acid–binding protein 4 (FABP4) in vascular endothelial cells. A and B, Gene (A) and protein (B) expression levels of FABP 4 in human coronary artery endothelial cells ( HCAECs ) treated with 50 ng/mL of vascular endothelial growth factor ( VEGF ) for 24 hours or 100 μmol/L H 2 O 2 for 1 hour followed by 23‐hour incubation in normal culture media after washing were determined by quantitative real‐time polymerase chain reaction (n=3 in each group) and Western blot analysis, respectively. * P <0.05 vs control. C, Experimental design of adenovirus‐mediated overexpression in HCAECs using adenovirus vector of FABP 4 (Ad‐ FABP 4) or empty sequence (Ad‐Control) ( HCAEC ‐ OE ). D, Gene expression of FABP 4 in HCAEC ‐ OE (n=6 in each group). * P <0.05 vs Ad‐Control. E, Representative Western blot analysis of FABP 4 in HCAEC ‐ OE . F, Western blot analysis of nitric oxide synthase ( NOS 3) and phosphorylated NOS 3 ( pS 1177) in HCAEC ‐ OE treated with 0.5 μmol/L insulin or 50 ng/mL VEGF for 0.5 hours (n=3 in each group). * P <0.05. G and H, Gene expression of inflammatory cytokines (G) and adhesion‐related molecules (H) in HCAEC ‐ OE (n=6 in each group). * P <0.05 vs Ad‐Control. I, Western blot analysis of FABP 4 (exposure: light, L; dark, D) and GAPDH was performed using the cell lysate ( CL ) and conditioned medium ( CM ) of HCAECs supplemented with 0.5% BSA in the absence and presence of 10 μmol/L isoproterenol for 6 to 24 hours (n=3 in each group). * P <0.05. J, Secretion of FABP 4 for 24 hours in HCAEC ‐ OE measured using an enzyme‐linked immunosorbent assay kit (n=3 in each group). Values were normalized to total protein concentration of the cell lysate. * P <0.05 vs Ad‐Control. AU indicates arbitrary unit; Icam1, including intracellular adhesion; IL, interleukin; Itga5, integrin a 5; Itgb3, integrin b 3; Mcp1, monocyte chemotactic protein‐1; Sele, selectin E; Tnfa, tumor necrosis factor α.

Article Snippet: Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza.

Techniques: Over Expression, Binding Assay, Expressing, Incubation, Real-time Polymerase Chain Reaction, Western Blot, Plasmid Preparation, Sequencing, Enzyme-linked Immunosorbent Assay, Protein Concentration

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Ectopic Fatty Acid–Binding Protein 4 Expression in the Vascular Endothelium is Involved in Neointima Formation After Vascular Injury

doi: 10.1161/JAHA.117.006377

Figure Lengend Snippet: Effects of the conditioned medium of Fabp4 ‐overexpressed endothelial cells in vascular smooth muscle cells. A, Experimental design of human coronary artery smooth muscle cells ( HCASMCs ) treated with the conditioned medium ( CM ) prepared by 24‐hour incubation of adenovirus vector of fatty acid–binding protein 4 (Ad‐ FABP 4)– and empty sequence (Ad‐Control)–transfected human coronary artery endothelial cells ( HCAECs ) in the absence and presence of 10 μg/mL anti‐ FABP 4 antibody ( FABP 4‐Ab) for 24 hours ( HCASMC ‐ CM ‐Ab). B, Gene expression of inflammatory cytokines and proliferation‐ and adhesion‐related molecules determined by quantitative real‐time polymerase chain reaction (PCR) in HCASMC ‐ CM ‐Ab (n=6 in each group). * P <0.05 vs CM of Ad‐Control–transfected HCAECs ( CM ‐Ad‐Control). † P <0.05 vs CM of Ad‐ FABP 4–transfected HCAECs ( CM ‐Ad‐ FABP 4). C and D, Proliferation of HCASMC ‐ CM supplemented with 5% FBS in the absence and presence of 10 μg/mL FABP 4‐Ab for 24 hours assessed by MTS (C) and bromodeoxyuridine (BrdU; D) assays (n=6 in each group). * P <0.05 vs CM ‐Ad‐Control. † P <0.05 vs CM ‐Ad‐ FABP 4. E, Migration of HCASMC ‐ CM supplemented with 0.5% BSA in the absence and presence of 10 μg/mL FABP 4‐Ab for 15 hours assessed by scratch wound assay (n=6 in each group). * P <0.05 vs CM ‐Ad‐Control. † P <0.05 vs CM ‐Ad‐ FABP 4. F, Experimental design of HCASMCs coincubated with Ad‐ FABP 4– and Ad‐Control–transfected HCAECs using insert transparent wells for 24 hours ( HCASMC ‐ TW ). G and H, Gene expression of inflammatory cytokines (G) and proliferation‐ and adhesion‐related molecules (H) determined by quantitative real‐time PCR in HCASMC ‐ TW (n=6 in each group). * P <0.05 vs coincubation using transparent wells with Ad‐Control–overexpressed HCAECs ( TW ‐Ad‐Control). AU indicates arbitrary unit; IL, interleukin; Itga5, integrin a 5; Itgb3, integrin b 3; Mcp1, monocyte chemotactic protein‐1; Pdgfra, platelet‐derived growth factor receptor α; Pdgfrb, platelet‐derived growth factor receptor β; Tnfa, tumor necrosis factor α.

Article Snippet: Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza.

Techniques: Incubation, Plasmid Preparation, Binding Assay, Sequencing, Transfection, Expressing, Real-time Polymerase Chain Reaction, Migration, Scratch Wound Assay Assay, Derivative Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Ectopic Fatty Acid–Binding Protein 4 Expression in the Vascular Endothelium is Involved in Neointima Formation After Vascular Injury

doi: 10.1161/JAHA.117.006377

Figure Lengend Snippet: Effects of the conditioned medium of Fabp4 ‐overexpressed endothelial cells in vascular endothelial cells. A, Experimental design of human coronary artery endothelial cells ( HCAECs ) treated with the conditioned medium ( CM ) prepared by 24‐hour incubation of adenovirus vector of fatty acid–binding protein 4 (Ad‐ FABP 4)– and empty sequence (Ad‐Control)–transfected HCAECs ( HCAEC ‐ CM ) in the absence and presence of 10 μg/mL anti‐ FABP 4 antibody ( FABP 4‐Ab) for 24 hours ( HCAEC ‐ CM ‐Ab). B, Gene expression of inflammatory cytokines and adhesion‐related molecules determined by quantitative real‐time polymerase chain reaction (PCR) in HCAEC ‐ CM (n=6 in each group). * P <0.05 vs CM of Ad‐Control–overexpressed HCAECs ( CM ‐Ad‐Control). C, Gene expression of inflammatory cytokines and adhesion‐related molecules determined by quantitative real‐time PCR in HCAEC ‐ CM ‐Ab (n=4 in each group). * P <0.05 vs FABP 4‐Ab (−). D, Experimental design of HCAECs coincubated with Ad‐ FABP 4– and Ad‐Control–transfected HCAECs using insert transparent wells for 24 hours ( HCAEC ‐ TW ). E and F, Gene expression of inflammatory cytokines (E) and adhesion‐related molecules (F) determined by quantitative real‐time PCR in HCAEC ‐ TW (n=6 in each group). * P <0.05 vs coincubation using transparent wells with Ad‐Control–overexpressed HCAECs ( TW ‐Ad‐Control). AU indicates arbitrary unit; Icam1, including intracellular adhesion; IL, interleukin; Itga5, integrin a 5; Itgb3, integrin b 3; Mcp1, monocyte chemotactic protein‐1; Sele, selectin E; Tnfa, tumor necrosis factor α.

Article Snippet: Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza.

Techniques: Incubation, Plasmid Preparation, Binding Assay, Sequencing, Transfection, Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Ectopic Fatty Acid–Binding Protein 4 Expression in the Vascular Endothelium is Involved in Neointima Formation After Vascular Injury

doi: 10.1161/JAHA.117.006377

Figure Lengend Snippet: Effects of exogenous fatty acid–binding protein 4 (FABP4) treatment in vascular endothelial cells. A, Experimental design of human coronary artery endothelial cells ( HCAECs ) treated with 200 nmol/L recombinant FABP 4 ( HCAEC ‐Rec) in the absence and presence of 10 μg/mL anti‐ FABP 4 antibody ( FABP 4‐Ab) for 24 hours ( HCAEC ‐Rec‐Ab). B, Gene expression of inflammatory cytokines and adhesion‐related molecules determined by quantitative real‐time polymerase chain reaction (PCR) in HCAEC ‐Rec (n=3 in each group). * P <0.05 vs Rec‐ FABP 4 (−). C, Gene expression of inflammatory cytokines and adhesion‐related molecules determined by quantitative real‐time PCR in HCAEC ‐Rec‐Ab (n=3 in each group). * P <0.05 vs FABP 4‐Ab (−). D, Representative Western blot analysis of endogenous FABP 4, His‐tagged FABP 4 (His‐ FABP 4), and GAPDH was performed using the cell lysate ( CL ) and conditioned medium ( CM) of HCAECs treated with 0 to 200 nmol/L His‐tagged Rec‐ FABP 4 for 6 hours. E, Western blot analysis of nitric oxide synthase 3 ( NOS 3) phosphorylated NOS 3 ( pS 1177) in HCAEC ‐Rec‐Ab followed by stimulation with 50 ng/mL vascular endothelial growth factor ( VEGF ) for 0.5 hours (n=3 in each group). * P <0.05. AU indicates arbitrary unit; Icam1, including intracellular adhesion; IL, interleukin; Itga5, integrin a 5; Itgb3, integrin b 3; Mcp1, monocyte chemotactic protein‐1; Sele, selectin E; Tnfa, tumor necrosis factor α.

Article Snippet: Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza.

Techniques: Binding Assay, Recombinant, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular imaging and biology

Article Title: 2 nd Window NIR Imaging of Radiation Injury Mitigation Provided by Reduced Notch-Dll4 Expression on Vasculature

doi: 10.1007/s11307-023-01840-7

Figure Lengend Snippet: Study design and number of animals for each experiment

Article Snippet: About ten million lung cells per animal were used for CD31 + enrichment using the CD31 Microbeads kit (cat# 130–109–680, Miltenyi) as per the manufacturer’s instructions.

Techniques: In Vivo, Imaging, Ex Vivo, Expressing, Immunostaining

Journal: Molecular imaging and biology

Article Title: 2 nd Window NIR Imaging of Radiation Injury Mitigation Provided by Reduced Notch-Dll4 Expression on Vasculature

doi: 10.1007/s11307-023-01840-7

Figure Lengend Snippet: SS.BN3 rats have reduced Dll4 expression and reduced susceptibility to late radiation-induced morbidity. a Consomic map illustrating the SS.BN3 rat is generated by the substitution of chromosome 3 from the Brown Norway rat into the Dahl-SS genetic background. b mRNA expression of Dll4 from CD31+ endothelial cells in non-irradiated SS (Dll4-high) and consomic SS.BN3 (Dll4-low) rats (n = 4 (SS) or 5 (SS.BN3) rats per group). c Survival following 13 Gy partial body irradiation (PBI) in adult SS (n = 13) and SS.BN3 rats (n = 19). P value for log-rank analysis: P = 0.0002 for SS vs SS.BN3 following 13 Gy PBI. d Change in body weight relative to pre-irradiation body weight for the rats represented in the survival curve (**P = 0.0066, ****P < 0.0001, two-way ANOVA Sidak’s multiple testing correction). Note the increased drop in body weight at 90 and 120 days in SS as compared to SS.BN3 rats, that occurs during pneumonitis and nephropathy, respectively

Article Snippet: About ten million lung cells per animal were used for CD31 + enrichment using the CD31 Microbeads kit (cat# 130–109–680, Miltenyi) as per the manufacturer’s instructions.

Techniques: Expressing, Generated, Irradiation

Journal: Molecular imaging and biology

Article Title: 2 nd Window NIR Imaging of Radiation Injury Mitigation Provided by Reduced Notch-Dll4 Expression on Vasculature

doi: 10.1007/s11307-023-01840-7

Figure Lengend Snippet: SS.BN3 (Dll4-low) rats have reduced radiation-induced loss of CD31+ endothelial cells. a Left, representative FACS contour plots showing the percentage of CD45−CD31+ lung ECs at day 70 following 13 Gy partial body irradiation in SS (Dll4-high) and SS.BN3 (Dll4-low) rats. Right, quantification of percent CD45−CD31+ cells (n = 9 rats per group). b Left, representative FACS contour plots showing the percentage of CD45−CD31+ kidney ECs at day 70 following 13 Gy partial body irradiation in SS (Dll4-high, n = 4) and SS.BN3 (Dll4-low, n = 5) rats. Right, quantification of percent CD45−CD31+ cells

Article Snippet: About ten million lung cells per animal were used for CD31 + enrichment using the CD31 Microbeads kit (cat# 130–109–680, Miltenyi) as per the manufacturer’s instructions.

Techniques: Irradiation