Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 1. Effect of angiotensin-II (Ang-II) on arteriolar leukocyte adhesion (A), venular leukocyte adhesion (B), and venular leu kocyte emigration (C) on mouse cremasteric microcirculation. CX3CL1 was expressed in the cremasteric arterioles and postcapil lary venules of Ang-II–injected animals (D). Results are mean±SEM of n=4 to 6 animals per group. **P<0.01 relative to vehicle-treated mice; +P<0.05 relative to CX3CR1−/+-treated mice.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Injection

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 2. Angiotensin-II (Ang-II) induces CX3CL1 mRNA (A and B) and protein (flow cytometry; C and D) expression in human umbilical arterial endothelial cells (HUAEC) and human umbilical vein endothelial cells (HUVEC). A neutralizing antibody against CX3CL1 function inhibited the recruitment of mononuclear leukocytes to Ang-II–stimulated HUAEC but not HUVEC (E and F). Results are mean±SEM of n=4 to 6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 relative to values in the Ang-II group. CX3CL1 expression was visualized in nonpermeabilized HUAEC and HUVEC by immunofluorescence (green). Nuclei were coun terstained with 4′,6-diamidino-2-phenylindole. Results are representative of n=5 independent experiments. In the flow chamber assay, results are mean±SEM of n= 5 to 7 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 rela tive to Ang-II+MOPC21 group. TNFα indicates tumor necrosis factor-α.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Flow Cytometry, Expressing, Immunofluorescence, Boyden Chamber Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 4. Angiotensin-II (Ang-II)–induced CX3CL1 expression (A) and mononuclear cell arrest (B) are inhibited by apocynin but not by allopurinol. Nox 5 but not Nox 2 or Nox 4 small interfering RNA (siRNA) inhibits Ang-II–induced CX3CL1 expression and mononuclear cell arrest in human umbilical arterial endothelial cells (HUAEC; C–H). HUAEC were stimulated with 1 µmol/L Ang-II for 24 hours. Some cells were pretreated with apocynine (30 μmol/L) or allopurinol (100 μmol/L) 1 hour before Ang-II stimulation. Results are the mean±SEM of n=7 independent experiments. **P<0.01 relative to values in the medium group; ++P<0.01 relative to values in the Ang-II–stimulated group. In endothelial cells transfected with Nox 2, Nox 4 or Nox 5 siRNA or control siRNA, results are mean±SEM of n=6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to values in Ang-II group in control siRNA–transfected cells. XO indicates xantine oxidase.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Small Interfering RNA, Transfection, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 3. Tumor necrosis factor-α (TNFα) mRNA (A) and pro tein (B) expression in human umbilical arterial endothelial cells (HUAEC) is abolished in HUAEC transfected with small interfer ing RNA (siRNA) targeting TNFα. TNFα siRNA inhibits Ang-II– induced CX3CL1 expression (C) and mononuclear–endothelial cell interactions (D) in HUAEC. Results are mean±SEM of n= 6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to their respective group in control siRNA–transfected cells. MFI indicates mean fluorescence intensity.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Transfection, Control, Fluorescence

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 6. Combined stimulation with tumor necrosis factor-α (TNFα), interferon γ (IFNγ), and angiotensin-II (Ang-II) promotes increased CX3CL1 expression (A) and mononuclear–endothelial cell interactions (B). Cells were treated with Ang-II (1 µmol/L), TNFα (20 ng/mL), IFNγ (20 ng/mL), Ang-II+TNFα, Ang-II+IFNγ, TNFα+IFNγ, or Ang-II+TNFα+IFNγ for 24 hours. Results are the mean±SEM of n=6 to 9 independent experiments (A) and n=6 to 7 independent experiments (B). *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to val ues in the Ang-II group; &P<0.05 or &&P<0.01 relative to values in the TNFα group; ##P<0.01 relative to values in the IFNγ group.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways.

doi: 10.2147/jir.s4019

Figure Lengend Snippet: Figure 2 Cell surface expression and mRNA expression of CX3CL1 in RA OBs. (A) Representative photomicrographs showing the immunohistochemical localization of antigenic CX3CL1 in RA OBs stimulated with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL) for 24 h (original magnifi cation, 400×). (B) Flow cytometric analysis of the level of CXC3L1 expression. Cultured OBs from RA patients were incubated for 24 h with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL). (C) Real-time RT-PCR analysis of CX3CL1 mRNA expression in RA OBs. Cultured OBs from RA patients were incubated for 4 h with TNF-α (20 ng/mL) and IFN-γ (1000 U/mL). Data are means ± SEM of 3 independent experiments. *p 0.05 vs control medium. (D) Representative agarose gel analysis of CX3CL1 semiquantitative RT-PCR products.

Article Snippet: OBs were labeled with monoclonal mouse PE-conjugated anti-CX3CL1 Ab (R and D Systems).

Techniques: Expressing, Immunohistochemical staining, Cell Culture, Incubation, Quantitative RT-PCR, Control, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways.

doi: 10.2147/jir.s4019

Figure Lengend Snippet: Figure 4 NF-κB expression in OBs and its role in the induction of CX3CL1 expression. (A) Real-time RT-PCR analysis revealed enhanced expression of NF-κB mRNA in RA OBs incubated for 4 h with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL). (B and C) Effects of NF-κB-specifi c siRNA on CX3CL1 secretion and mRNA expression. RA OBs transfected with either NF-κB-specifi c siRNA or negative control siRNA were incubated for 24 h (B) or 4 h (C) with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL), after which culture supernatants (B) or total RNA (C) were isolated and ELISA or real-time PCR for CX3CL1 were conducted. Data are means ± SEM from 3 independent experiments.

Article Snippet: OBs were labeled with monoclonal mouse PE-conjugated anti-CX3CL1 Ab (R and D Systems).

Techniques: Expressing, Quantitative RT-PCR, Incubation, Transfection, Negative Control, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of inflammation research

Article Title: Synergistic induction of CX3CL1 by TNF alpha and IFN gamma in osteoblasts from rheumatoid arthritis: involvement of NF-kappa B and STAT-1 signaling pathways.

doi: 10.2147/jir.s4019

Figure Lengend Snippet: Figure 6 STAT-1 expression in OBs and its role in the induction of CX3CL1 expression. (A) Real-time RT-PCR analysis revealed enhanced expression of STAT-1 mRNA in RA OBs incubated for 4 h with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL) and in RA OBs incubated with IFN-γ alone. (B and C) Effects of STAT-1-specifi c siRNA on CX3CL1 secretion and expression. RA OBs transfected with either STAT-1-specifi c siRNA or negative control siRNA were incubated for 24 h (B) or 4 h (C) with TNF-α (20 ng/mL) plus IFN-γ (1000 U/mL), after which culture supernatants (B) or total RNA (C) was isolated and ELISA or real-time PCR for CX3CL1 was conducted. Data are means ± SEM from three independent experiments. *p 0.05 vs medium alone (A), *p 0.05 vs TNF-α/IFN-γ with control siRNA (B and C).

Article Snippet: OBs were labeled with monoclonal mouse PE-conjugated anti-CX3CL1 Ab (R and D Systems).

Techniques: Expressing, Quantitative RT-PCR, Incubation, Transfection, Negative Control, Isolation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Control