Journal: Circulation

Article Title: Enhanced Inhibition of Neointimal Hyperplasia by Genetically Engineered Endothelial Progenitor Cells

doi: 10.1161/01.cir.0000121732.85572.6f

Figure Lengend Snippet: Figure 1. Characterization of rabbit EPCs and comparison with HUVECs. A, B, Cobblestone morphology of EPCs and HUVEC 10 days after plating (40); C, D, immunohistochemical detec- tion of CD31; immunofluorescence detection of (E, F), VE-cadherin; (G, H), eNOS; (I, J), vWF. Preparations were viewed at 400 unless otherwise indicated. Preparations in G, H, I, and J were counterstained with Hoechst 33342.

Article Snippet: The cells were incubated with 1:50 dilution of monoclonal anti human CD31 (PECAM-1, Dako, Carpinteria, Calif), or polyclonal goat anti-mouse VE-cadherin or anti-human eNOS (R&D Systems Inc, Minneapolis, Minn), or anti-human von Willebrand factor (vWF) (Dako).

Techniques: Comparison, Immunohistochemical staining

Journal: Circulation

Article Title: Enhanced Inhibition of Neointimal Hyperplasia by Genetically Engineered Endothelial Progenitor Cells

doi: 10.1161/01.cir.0000121732.85572.6f

Figure Lengend Snippet: Figure 2. Ex vivo retroviral transduction and transgene expression in EPCs. A, Confluent monolayer of EPCs trans- duced with MSCV retrovirus expressing GFP viewed under normal (white) light (100); B, view of same field under green fluorescent light; C, D, human eNOS transgene mRNA and protein expression in untransduced and trans- duced EPCs, respectively; E, F, HO-1 mRNA and protein expression in untransduced and transduced EPCs, respectively.

Article Snippet: The cells were incubated with 1:50 dilution of monoclonal anti human CD31 (PECAM-1, Dako, Carpinteria, Calif), or polyclonal goat anti-mouse VE-cadherin or anti-human eNOS (R&D Systems Inc, Minneapolis, Minn), or anti-human von Willebrand factor (vWF) (Dako).

Techniques: Ex Vivo, Retroviral, Transduction, Expressing

Journal: Circulation

Article Title: Enhanced Inhibition of Neointimal Hyperplasia by Genetically Engineered Endothelial Progenitor Cells

doi: 10.1161/01.cir.0000121732.85572.6f

Figure Lengend Snippet: Figure 3. In vivo transgene expression after transplantation of EPCs onto injured carotid arteries. A, Immunohistochemical detection of eNOS (arrows) in frozen sections from denuded vessel trans- planted with eNOS-EPCs (250). B, eNOS immunoreactivity in sections from control vessel transplanted with GFP- EPCs (250); C, immunohistochemical detection of human HO-1 (arrows) in fro- zen sections from vessel transplanted with HO-1-EPCs (250); D, control ves- sels transplanted with GFP-EPCs (250); E, detection of human eNOS transgene mRNA expression by RT-PCR; F, human eNOS protein expression in homoge- nates from eNOS-EPC–transplanted ves- sels; G, human HO-1 transgene mRNA expression; H, HO-1 protein expression in homogenates from HO-1-EPC–trans- planted vessels. GFP-EPC–transplanted vessels served as controls.

Article Snippet: The cells were incubated with 1:50 dilution of monoclonal anti human CD31 (PECAM-1, Dako, Carpinteria, Calif), or polyclonal goat anti-mouse VE-cadherin or anti-human eNOS (R&D Systems Inc, Minneapolis, Minn), or anti-human von Willebrand factor (vWF) (Dako).

Techniques: In Vivo, Expressing, Transplantation Assay, Immunohistochemical staining, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Circulation

Article Title: Enhanced Inhibition of Neointimal Hyperplasia by Genetically Engineered Endothelial Progenitor Cells

doi: 10.1161/01.cir.0000121732.85572.6f

Figure Lengend Snippet: Figure 4. Reendothelialization of injured arteries after transplan- tation of EPCs expressing GFP, eNOS, or HO-1 genes. Low- power (100) magnification of vessel profiles stained with anti- CD31 antibody in A, unseeded; B, GFP-EPCs; C, eNOS-EPCs; and D, HO-1-EPCs. Red CD31-positive staining indicates pres- ence of an endothelial monolayer. E, Quantification of endotheli- alization by planar morphometry. Endothelialization was enhanced in EPC-transplanted vessels compared with unseeded saline-treated controls (*P0.05, n4/group).

Article Snippet: The cells were incubated with 1:50 dilution of monoclonal anti human CD31 (PECAM-1, Dako, Carpinteria, Calif), or polyclonal goat anti-mouse VE-cadherin or anti-human eNOS (R&D Systems Inc, Minneapolis, Minn), or anti-human von Willebrand factor (vWF) (Dako).

Techniques: Expressing, Staining, Saline

Journal: Circulation

Article Title: Enhanced Inhibition of Neointimal Hyperplasia by Genetically Engineered Endothelial Progenitor Cells

doi: 10.1161/01.cir.0000121732.85572.6f

Figure Lengend Snippet: Figure 5. Inhibition of neointimal proliferation by EPCs in injured carotid arteries. All sections were stained with Accustain elastic stain. Low (40, left) and high (200, right) magnification of vessel profiles. A, B, Saline-injured artery transplanted with eNOS-EPCs (n6); C, D, balloon-injured artery transplanted with EPC expressing GFP (n6); E, F, balloon-injured artery transplanted with EPC expressing eNOS (n6); G, H, balloon- injured artery transplanted with HO-1-EPCs (n6); I, neointima/ media ratios in unseeded and EPC-transplanted injured vessels. Arrows indicate neointima. *Saline vs GFP-EPCs, eNOS-EPCs, HO-1-EPCs; †eNOS-EPCs vs GFP-EPCs, P0.05.

Article Snippet: The cells were incubated with 1:50 dilution of monoclonal anti human CD31 (PECAM-1, Dako, Carpinteria, Calif), or polyclonal goat anti-mouse VE-cadherin or anti-human eNOS (R&D Systems Inc, Minneapolis, Minn), or anti-human von Willebrand factor (vWF) (Dako).

Techniques: Inhibition, Staining, Saline, Expressing

Journal: Nutrients

Article Title: Analysis of the Combined Effects of a Novel Combination of Hypersmin, Pumpkin Seed and Amaranthus Extracts in an In Vitro Model of Chronic Venous Insufficiency.

doi: 10.3390/nu17111807

Figure Lengend Snippet: Figure 3. Vascular tone analysis on in vitro model of vein after intestinal passage. In (A) NO production; in (B) eNOS levels; in (C) endothelin-1 levels. From (A) to (C) all the data are obtained with specific ELISA kits; * p < 0.05 vs. Control; α p < 0.05 vs. single agents; β p < 0.05 vs. Commercial product; γ p < 0.05 vs. KCl.

Article Snippet: The eNOS concentration was measured following the manufacturer’s instructions for the DuoSet ELISA kit for Human eNOS (R&D Systems, Minneapolis, MN, USA) in HUVEC cells [39].

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of vascular surgery

Article Title: Neointimal hyperplasia on a cell-seeded polytetrafluoroethylene graft is promoted by transfer of tissue plasminogen activator gene and inhibited by transfer of nitric oxide synthase gene.

doi: 10.1016/j.jvs.2004.10.023

Figure Lengend Snippet: Fig. 2. Immunostaining of neointima. Paraffin sections of grafts seeded with SMC/tPA (a), SMC/tPANOS (c), or SMC (b, d) were stained with antibody against human tPA (a, b) or antibody against human eNOS (c, d). Black arrows indicate positively stained cells in brown. White arrows indicate negtively stained cells with nuclear stained in blue.

Article Snippet: Biotinylated anti-human eNOS goat immunoglobulin G (1:5 dilution) was from R & D Systems, Inc (Minneapolis, Minn).

Techniques: Immunostaining, Staining

Journal: Current Research in Physiology

Article Title: The comparison of endothelial function of moderate intensity interval exercise with continuous exercise in healthy men

doi: 10.1016/j.crphys.2022.07.003

Figure Lengend Snippet: Comparison of vascular markers between intermittent and continuous exercise.

Article Snippet: Then, the enzyme-linked immunosorbent assay (ELISA) method was used for determination of serum concentrations of N-terminal proANP (NTproANP), N-terminal proBNP (NTproBNP), N-terminal proCNP (NTproCNP), endothelial nitric oxide synthase activity, endothelin 1 (ET-1), adiponectin, and leptin (Elabscience Cat. No: E-EL-H1848, Cat. No: E-EL-H0902, Cat. No: E-EL-H2538, Cat. No: E-EL-H0755, Cat. No: E-EL-H0064, Cat. No: E-EL-H0004, Cat. No: E-EL-H0113 respectively).

Techniques: Comparison, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: Transcriptomic changes upon NANOS1 and NANOS3 overexpression in TCam-2 cell line. ( A ) RNA-Sequencing (RNA-Seq) upon NANOS1 (left panel) and NANOS3 (right panel) overexpression visualized as MA-plots. Differentially expressed genes were filtered with log2FC ≥ 0.5 for upregulated and ≤ −0.5 for downregulated genes. Adjusted p -value ≤ 0.01 was considered as significant. ( B ) Venn diagram showing common and distinct differentially expressed genes between NANOS1 and NANOS3 overexpression. ( C ) Expression level of infertility genes downregulated upon NANOS1 (upper heatmap) and NANOS3 (lower heatmap) overexpression visualized as heatmaps with z-score scaling. ( D ) Cancer-germ cell genes downregulated upon NANOS1 (upper heatmap) and NANOS3 (lower heatmap) overexpression. Genes involved in cell cycle are marked by arrows.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: Over Expression, RNA Sequencing Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: Biological clustering analysis (BCA) reveals cell cycle related functions for NANOS1 and NANOS3. ( A ) Workflow for biological clustering analysis (BCA). High confidence (score ≥ 0.9) protein–protein interaction networks of differentially expressed genes are created using the STRING database followed by cluster detection by MCODE. Further, these clusters were subjected to gene ontology (GO) search for the corresponding biological process. ( B ) NANOS1 and ( C ) NANOS3 downregulated cell cycle cluster containing infertility (shown in black diamonds, ) and cancer-germ cell genes (shown in black triangles, ).

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: Distinct phenotypic effects of NANOS1 and NANOS3 overexpression on the cell cycle. ( A ) Cell cycle related biological processes downregulated upon NANOS1 (top panel) and NANOS3 (bottom panel) overexpression identified in GO analysis. ( B ) Cell cycle analysis by propidium iodide staining followed by flow cytometry analysis upon NANOS1 (top panel) and NANOS3 (bottom panel) overexpression. ( C ) Apoptosis analysis by Annexin-V staining followed by flow cytometry analysis upon NANOS1 and NANOS3 overexpression. Standard error is visualized as error bars. * = p -value ≤ 0.05.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: Over Expression, Cell Cycle Assay, Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: FOXM1 and TAF1 act as transcription factors for NANOS1 and NANOS3 downregulated genes. ( A ) Cumulative distribution analysis of differentially expressed genes upon NANOS3 RNA-Seq. Common differentially expressed mRNA distribution shown in blue and labeled as targets. The rest of the differentially expressed genes are shown in red and labeled as non-targets. ( B ) Common differentially expressed genes that are repressed by both NANOS3 and PUM1 identified as downregulated (log2FC < 0) upon NANOS3 overexpression and upregulated (log2FC > 0) upon PUM1 knockdown. ( C ) iRegulon transcription factor prediction analysis for NANOS3/PUM1 common repressed genes. ( D ) Cell cycle related biological processes identified by gene ontology enrichment analysis for NANOS3-PUM1 common repressed genes.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: RNA Sequencing Assay, Labeling, Over Expression

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: FOXM1 mRNA is regulated by the NANOS3/PUM1 post-transcriptional complex through its’ 3′UTR. ( A ) Western-blot visualization of GST pull-down of NANOS3 with and without RNase A. ( B ) Schematic representation of FOXM1 3′UTR containing the Pumilio binding element (PBE-UGUAHAUA) and 2 PBE-like motifs (containing core UGUA motif). ( C ) Dual luciferase assay to measure Renilla/Firefly ratio upon PUM1 knockdown and NANOS3 overexpression using 3′UTR of FOXM1 in the downstream of Renilla ORF. Standard error is visualized as error bars. * = p -value ≤ 0.05, *** = p -value ≤ 0.001.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: Western Blot, Binding Assay, Luciferase, Over Expression

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: The NANOS3–PUM1–FOXM1 axis coordinates the expression of the cell cycle genes. ( A ) Dual luciferase assay to measure Renilla/Firefly ratio upon PUM1 knockdown and NANOS3 overexpression using 3′UTR of CCNA2 , KIF20B , and RAD21 in the downstream of Renilla ORF. ( B ) Relative fold change of FOXM1 binding events per 1000 cells for the promoter region of the target genes measured by ChIP-qPCR. ( C ) Differential correlation analysis of NANOS3 , PUM1 , and FOXM1 in publicly available healthy testis (GTEx) and testicular cancer datasets (TCGA). Each dot represents a single GTEx or TCGA dataset. Expression values are transformed as log(expression +1). Standard error is visualized as error bars. ** = p -value ≤ 0.01, *** = p -value ≤ 0.001.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: Expressing, Luciferase, Over Expression, Binding Assay, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model

doi: 10.3390/ijms23126592

Figure Lengend Snippet: Model of NANOS3-PUM1-FOXM1 axis regulating G2/M phase of the cell cycle in TCam-2 cell line. Created with BioRender.com.

Article Snippet: Transient overexpression was performed by transfecting cells with pCMV6-entry vector (OriGene) containing NANOS1 (#RC219123), NANOS3 (#RC221867), and PUM1 (#RC201219) ORF tagged with FLAG (DYKDDDDK) peptide in the C-terminus.

Techniques: