2001 1 5  (Vector Laboratories)

Journal: International Journal of Physiology, Pathophysiology and Pharmacology

Article Title: Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury

doi:

Figure Lengend Snippet: Immunolocalization of Ref-1 in the liver. A: Immunostaining for Ref-1 alone (brown), with hematoxylin counterstain. The arrow indicates a hepatocyte nucleus positive for Ref-1; the arrowhead indicates a sinusoidal lining cell positive for Ref-1. Positive staining was also apparent in cells lining the bile duct (BD). B: Double staining for Ref-1 (black) and glutamine synthetase (GS; brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in hepatocytes; the arrow indicates a hepatocyte nucleus positive for Ref-1. HV: terminal hepatic venule C: Double staining for Ref-1 (black) and β-catenin (brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in biliary epithelial cells; the arrow indicates a hepatocyte nucleus positive for Ref-1. D: Double staining for Ref-1 (black) and Trx-1 (brown). Arrowheads indicate colocalization in bile ducts (BD); double arrows indicate colocalization in sinusoidal lining cells (SEC; Kupffer cells). The arrow indicates a hepatocyte positive for Ref-1. Sections in panels B-D were not counterstained. All panels are images from young, control rats and are representative of 5 individual animals. The cellular localization of Ref-1 was identical in old, nonheated rats (not shown). Scale bar = 100 μm.

Article Snippet: Sections were washed in PBS, then incubated in biotinylated secondary antibody at a 1:350 dilution (mouse for Ref-1 and rabbit for Trx-1; Vector Labs, BA-2001, and BA-1100, respectively) for one hour at room temperature, after which they were washed in PBS, then incubated in avidin-biotin complex (Vector Labs, PK-4000).

Techniques: Immunostaining, Staining, Double Staining, Expressing

Journal: International Journal of Physiology, Pathophysiology and Pharmacology

Article Title: Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury

doi:

Figure Lengend Snippet: Localization of hepatic Trx-1. A: Trx-1 staining in a young, control liver. An identical pattern of cellular localization was observed in the old control rats (not shown). The arrowhead indicates positive staining in a bile duct, arrows indicate positive staining in nonparenchymal cells. B: Colocalization of Trx-1 (black) and HO-1 (brown) in a young control rat; double-positive cells are indicated by arrowheads (results are representative of 3 separate young and old control animals, with the experiment run in triplicate). C: Colocalization of Trx-1 (brown) and iron deposits (cyan) in an old, control rat, double positive cells are indicated by arrowheads (results are representative of 3 separate old animals). Scale bar = 100 μm.

Article Snippet: Sections were washed in PBS, then incubated in biotinylated secondary antibody at a 1:350 dilution (mouse for Ref-1 and rabbit for Trx-1; Vector Labs, BA-2001, and BA-1100, respectively) for one hour at room temperature, after which they were washed in PBS, then incubated in avidin-biotin complex (Vector Labs, PK-4000).

Techniques: Staining

Journal: International Journal of Physiology, Pathophysiology and Pharmacology

Article Title: Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury

doi:

Figure Lengend Snippet: Increase in Trx-1 abundance in old animals after hyperthermia. Left panel: representative immunoblot of Trx-1 in young and old animals under control conditions, and at the indicated times after heat stress (in hours). Ponceau stained membrane (Pon) demonstrates equal loading and transfer. Right panel: quantitation of Trx-1 expression, normalized to the Ponceau stain and further normalized to the young, nonheated group. †Significant effect of heat stress (n = 7-9 young and old animals in the nonheated condition, and at each time point after heat stress).

Article Snippet: Sections were washed in PBS, then incubated in biotinylated secondary antibody at a 1:350 dilution (mouse for Ref-1 and rabbit for Trx-1; Vector Labs, BA-2001, and BA-1100, respectively) for one hour at room temperature, after which they were washed in PBS, then incubated in avidin-biotin complex (Vector Labs, PK-4000).

Techniques: Western Blot, Staining, Quantitation Assay, Expressing

Journal: International Journal of Physiology, Pathophysiology and Pharmacology

Article Title: Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury

doi:

Figure Lengend Snippet: Colocalization of Ref-1 and Trx-1 within ductular reactions of aged animals. A: Ductular reaction (DR) in an old rat showing positive staining for Ref-1. The arrow indicates a cell positive for Ref-1. Multiple nuclei within bile ducts cells are positive for Ref-1. B: Ductular reaction in an old rat showing positive staining for Trx-1. The arrow indicates a cell positive for Trx-1. Frequent positive staining for Trx-1 was observed in cells within DR. Sections in Panels A and B were counterstained with hematoxylin. C: Colocalization of Ref-1 (black) and Trx-1 (brown) in cells of a DR. Arrowheads indicate cells positive for both Ref-1 and Trx-1. D: Colocalization of Ref-1 (black) and β-catenin (brown) in biliary epithelial cells of a DR. Arrowheads indicate cells positive for both Ref-1 and β-catenin. Sections in Panels C and D were not counterstained. Scale bar = 100 μm.

Article Snippet: Sections were washed in PBS, then incubated in biotinylated secondary antibody at a 1:350 dilution (mouse for Ref-1 and rabbit for Trx-1; Vector Labs, BA-2001, and BA-1100, respectively) for one hour at room temperature, after which they were washed in PBS, then incubated in avidin-biotin complex (Vector Labs, PK-4000).

Techniques: Staining