Journal: Molecular neurobiology

Article Title: Autologous bone marrow mononuclear cell transplantation delays progression of carotid atherosclerosis in rabbits

doi: 10.1007/s12035-015-9347-3

Figure Lengend Snippet: BMMNC transplantation reverses atherosclerosis-induced changes in antioxidant enzymes, eNOS, inflammatory response, and cell apoptosis. (A) Western blot analysis of eNOS in plaques. (B) Quantification analysis showed that plaques from the BMMNC group contained more eNOS than did plaques from the vehicle group. (C–F) BMMNC transplantation upregulated expression of the antioxidant enzymes GPx-1 and SOD-1 in plaque. (C, E) Western blot analysis of GPx-1 and SOD-1 in plaque. (D, F) Quantification of band densities showed that plaques from rabbits treated with BMMNCs had significantly higher GPx-1 and SOD-1 protein expression than did plaques from vehicle-treated rabbits. (G–H) BMMNC transplantation reduced expression of cell-death marker cleaved caspase-3 in plaque. (G) Western blot analysis of cleaved caspase-3. (H) Quantification analysis showed that cleaved caspase-3 was significantly lower in plaques from BMMNC-treated rabbits than in plaques from vehicle-treated rabbits. *p<0.05 vs. vehicle group. (I–L) BMMNC transplantation significantly decreased the mRNA expression of proinflammatory cytokines in plaque. (I) Representative RT-PCR bands showing mRNA expression of IL-1 β, TNF-α, and MMP-9 in plaque. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (J–L) Analysis of band optical densities showed that BMMNC transplantation significantly decreased the mRNA expression of IL-1 β, TNF-α, and MMP-9 compared with that in vehicle-treated animals. *p<0.05 vs. vehicle group. n=6 rabbits per group.

Article Snippet: The primary antibodies were mouse anti-rabbit IGF-1 (1:200, Global Biological Technologies Inc., Shanghai, China), rabbit anti-rabbit IGF-1R (1:100, Biosynthesis), rabbit anti-rabbit eNOS (1:400, Boster), mouse anti-rabbit GPx-1 (1:100, Abcam), goat anti-rabbit SOD-1 (1:1000, Abcam), rabbit anti-rabbit caspase-3 (1:1000, Abcam), and mouse anti-rabbit β-actin (1:500, Boster).

Techniques: Transplantation Assay, Western Blot, Expressing, Marker, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Hypertension

Article Title: Renal Collecting Duct NOS1 Maintains Fluid–Electrolyte Homeostasis and Blood Pressure

doi: 10.1161/hypertensionaha.113.01291

Figure Lengend Snippet: Figure 1. NOS1 splice variant expression of the inner medulla (IM) and cerebellum (CB) from wild-type mouse (WT) and NOS1αKO mouse as determined by Western blotting with anti- NOS1 (C terminus). A, NOS1α is expressed in the cerebellum of the WT mouse but not in the cerebellum of NOS1αKO mice. NOS1β is expressed in the IM of WT and NOS1αKO mice. The molecular weight marker (MW) was run on the same gel, but it was noncontiguous. B, NOS1β and NOS3 coexpressed in the IM, and NOS1α and NOS3 coexpressed in the cerebellum of the WT mouse. The molecular weight marker (MW) was run on the same gel, but it was noncontiguous. C, Collecting duct expression of NOS1β in the WT and NOS1αKO mice.

Article Snippet: IHC - immunohistochemistry Antibody Sequence Amino Acids Host Application Concentration Company Location NOS1 rat C-terminus rabbit polyclonal IB 20 μg/10ml Santa Cruz Santa Cruz, CA NOS1 human 1414-1434 rabbit polyclonal IB, IHC 1.4 μg Biomol Plymouth Meeting, PA NOS3 human 1025-1203 Mouse monoclonal IB 12.5 μg/ 10ml BD Biosciences San Jose, CA CD3-ε mouse C-terminus goat polyclonal IHC 0.2 μg Santa Cruz Santa Cruz, CA F4/80 mouse ? rat monoclonal IHC 0.1 μg AbD Serotech Raleigh, NC 3 NOS1!"

Techniques: Variant Assay, Expressing, Western Blot, Molecular Weight, Marker

Journal: Redox Biology

Article Title: Reciprocal regulation of eNOS, H 2 S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin

doi: 10.1016/j.redox.2017.02.006

Figure Lengend Snippet: eNOS and Akt phosphorylation and expression correlate with atheroma stability. Representative western blots and relative densitometric analysis of Α. p-eNOS/ t -eNOS and t -eNOS/β-actin B. Immunohistochemical (IHC) analysis of eNOS in serial sections from stable (n=8) and unstable plaques (n=8). For IHC purposes, frames in 200x magnification pictures represent the areas of 400x magnification. C. p-Akt (S473)/ t -Akt and t -Akt/β-actin (*p<0.05 vs Unstable).

Article Snippet: a. Antibodies: For immunohistochemical analysis the following antibodies were used: anti-eNOS (6H 2 ) mouse monoclonal antibody at a 1:100 dilution (Cell Signaling Technology, Beverly, MA, USA), anti- NOX4 (IgG rabbit polyclonal; epitope: a synthetic peptide made to an internal region of the human NOX4 protein (between residues 100–200)) at a 1:50 dilution (Novus-Biologicals, Europe), anti iNOS (Ab-1) (IgG rabbit polyclonal; epitope: purified enzyme from mouse macrophages (RAW 264.7) cells) in dilution 1:100 (Cayman Chemicals, Lab Supplies Greece). b.

Techniques: Phospho-proteomics, Expressing, Western Blot, Immunohistochemical staining

Journal: Redox Biology

Article Title: Reciprocal regulation of eNOS, H 2 S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin

doi: 10.1016/j.redox.2017.02.006

Figure Lengend Snippet: Simvastatin induces eNOS expression and phosphorylation independently of Akt. Representative western blots and relative densitometric analysis of Α. p-eNOS/ t -eNOS and t -eNOS/β-actin B. Immunohistochemical (IHC) analysis of eNOS in serial sections from s/nost (n=8) and s/st (n=8) groups For IHC purposes, frames in 200x magnification pictures represent the areas of 400x magnification. C. p-Akt (S473)/ t -Akt and t -Akt/β-actin (*p<0.05 vs s/nost).

Article Snippet: a. Antibodies: For immunohistochemical analysis the following antibodies were used: anti-eNOS (6H 2 ) mouse monoclonal antibody at a 1:100 dilution (Cell Signaling Technology, Beverly, MA, USA), anti- NOX4 (IgG rabbit polyclonal; epitope: a synthetic peptide made to an internal region of the human NOX4 protein (between residues 100–200)) at a 1:50 dilution (Novus-Biologicals, Europe), anti iNOS (Ab-1) (IgG rabbit polyclonal; epitope: purified enzyme from mouse macrophages (RAW 264.7) cells) in dilution 1:100 (Cayman Chemicals, Lab Supplies Greece). b.

Techniques: Expressing, Phospho-proteomics, Western Blot, Immunohistochemical staining

Journal: Redox Biology

Article Title: Reciprocal regulation of eNOS, H 2 S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin

doi: 10.1016/j.redox.2017.02.006

Figure Lengend Snippet: Summary of biochemical changes in atheromas.

Article Snippet: a. Antibodies: For immunohistochemical analysis the following antibodies were used: anti-eNOS (6H 2 ) mouse monoclonal antibody at a 1:100 dilution (Cell Signaling Technology, Beverly, MA, USA), anti- NOX4 (IgG rabbit polyclonal; epitope: a synthetic peptide made to an internal region of the human NOX4 protein (between residues 100–200)) at a 1:50 dilution (Novus-Biologicals, Europe), anti iNOS (Ab-1) (IgG rabbit polyclonal; epitope: purified enzyme from mouse macrophages (RAW 264.7) cells) in dilution 1:100 (Cayman Chemicals, Lab Supplies Greece). b.

Techniques:

Journal: Redox Biology

Article Title: Reciprocal regulation of eNOS, H 2 S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin

doi: 10.1016/j.redox.2017.02.006

Figure Lengend Snippet: Reciprocal regulation of eNOS vs H 2 S in atherosclerotic plaques. eNOS-derived NO increases bioavailability of NO in stable plaques, which would be expected to occur due to eNOS upregulation and phosphorylation (due to Akt activation); concomitantly iNOS and NOX-4 are downregulated presumably due to reduced HIF-1α. Moreover CBS and CSE positively correlate with plaque vulnerability. Simvastatin administration decreases HIF-1α expression and its downstream targets, iNOS and HO-1, whereas it increases the phosphorylation and expression of eNOS. Reduced CSE expression is associated with simvastatin administration and enhanced plaque stability (Modified by ). Red color indicates signaling molecules that decreased, whereas green color indicates signaling molecules that are upregulated.

Article Snippet: a. Antibodies: For immunohistochemical analysis the following antibodies were used: anti-eNOS (6H 2 ) mouse monoclonal antibody at a 1:100 dilution (Cell Signaling Technology, Beverly, MA, USA), anti- NOX4 (IgG rabbit polyclonal; epitope: a synthetic peptide made to an internal region of the human NOX4 protein (between residues 100–200)) at a 1:50 dilution (Novus-Biologicals, Europe), anti iNOS (Ab-1) (IgG rabbit polyclonal; epitope: purified enzyme from mouse macrophages (RAW 264.7) cells) in dilution 1:100 (Cayman Chemicals, Lab Supplies Greece). b.

Techniques: Derivative Assay, Phospho-proteomics, Activation Assay, Expressing, Modification