Journal: Aging (Albany NY)

Article Title: Voluntary aerobic exercise increases arterial resilience and mitochondrial health with aging in mice

doi: 10.18632/aging.101099

Figure Lengend Snippet: Aortic protein expression of PGC-1α ( A ), SIRT-3 ( B ), Fis1 ( C ), and Mfn2 ( D ) in arteries from young control (YC), old control (OC), young voluntary wheel running (YVR) and old voluntary wheel running (OVR) mice. Representative images are presented below each panel with corresponding images of normalizer (beta actin) taken from the same region of the same blot. Data are presented normalized to beta actin and relative to the mean of the YC group as means with error bars representing SEM, n=6-8/group. * p<0.05 vs. YC, # p=0.1 vs. YC.

Article Snippet: Proteins were transferred onto nitrocellulose membranes (Trans-Blot Turbo System, Bio-Rad Laboratories, Inc., Hercules, CA, USA) and incubated overnight with the following primary antibodies: peroxisome proliferator-activated receptor gamma co-activator 1-alpha (PGC-1α, 1:1000, Novus Biologicals, USA), NAD-dependent deacetylase sirtuin-3 (SIRT-3, 1:500, AbCam, Inc.), catalase (1:1000, AbCam, Inc.) heat shock protein 90 (Hsp90, 1:1000, Enzo Life Sciences, Inc.), Total OXPHOS Rodent Antibody Cocktail (containing antibodies against Complex I subunit NDUFB8, Complex II –30k, Complex III Core protein 2, Complex IV subunit I, and Complex V alpha subunit; 1:250, Novus Biologicals, USA), Fis1 (TTC11; 1:500, Novus Biologicals, USA), Mitofusin 2 (Mfn2; 1:500, AbCam, Inc.), and beta actin (normalizer, 1:2000, Cell Signaling Technology, Inc.).

Techniques: Expressing, Control

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Epigallocatechin-3-O-gallate decreases tumor necrosis factor-alpha-induced fractalkine expression in endothelial cells by suppressing NF-kappaB.

doi: 10.1159/000257494

Figure Lengend Snippet: Fig. 3. Immunoblot analysis of fractalkine protein expression in HUVECs after silencing NF-κB p65. HUVECs were transfected with NF-κB p65 siRNA or control siRNA (Cont siRNA) and treated with TNF-α with or without EGCG (50 µmol/L) for 6 h. Actin was used as the loading control. Densitometric analyses are presented as the relative ratio of fractalkine to actin. The relative ratio compared to the control is arbitrarily defined as 1. Note that silencing NF-κB p65 reduced TNF-α-induced fractalkine protein expression. Bars represent the mean±SD from 3 different experiments. *, p<0.05 vs. Cont siRNA only; #, p<0.05 vs. TNF-α plus Cont siRNA.

Article Snippet: Fractalkine-specific siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NF-κB p65 siRNA (human specific) (Cell Signaling Technology), negative control siRNA (Dharmacon Research, Lafayette, CO, USA) were used.

Techniques: Western Blot, Expressing, Transfection, Control

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Epigallocatechin-3-O-gallate decreases tumor necrosis factor-alpha-induced fractalkine expression in endothelial cells by suppressing NF-kappaB.

doi: 10.1159/000257494

Figure Lengend Snippet: Fig. 6. EGCG reduces adhesion of monocytes to HUVEC monolayers. (A) Representative images of the adhesion of monocytes to HUVEC monolayers. HUVECs were incubated with the indicated doses of EGCG, fractalkine-specific siRNA, control siRNA, antibody against fractalkine, control antibody (Cont antibody) and TNF-α (10 ng/mL) for 4 h. Fluorescence-labelled monocytes were added to the HUVEC monolayer and allowed to adhere for 2 h. The adhered cells were measured as described in Materials and Methods. (B) Quantification of monocyte adhesion to HUVECs. Data are from three experiments and are expressed as a percentage of adhesion to TNF-α-induced HUVECs. Bars represent the mean±SD from three experiments. *, p<0.05 vs. control (CB); #, p<0.05 vs. TNF-α.

Article Snippet: Fractalkine-specific siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NF-κB p65 siRNA (human specific) (Cell Signaling Technology), negative control siRNA (Dharmacon Research, Lafayette, CO, USA) were used.

Techniques: Incubation, Control, Fluorescence