nrf2 transcription factor assay kit (cat. no. 600590)  (Cayman Chemical)

Journal: Nutrients

Article Title: Accelerated Aging Induced by an Unhealthy High-Fat Diet: Initial Evidence for the Role of Nrf2 Deficiency and Impaired Stress Resilience in Cellular Senescence

doi: 10.3390/nu16070952

Figure Lengend Snippet: Nrf2 gene expression and DNA binding activity in Nrf2 +/+ and Nrf2 +/− mice. ( A ): Nrf2 mRNA levels in the cortex of Nrf2 +/+ and Nrf2 +/− mice ( n = 3–5). ( B , C ): DNA binding ability of Nrf2 assessed in the nuclear extracts of the cortex and liver of Nrf2 +/+ ( n = 4) and Nrf2 +/− ( n = 4–5) mice using Trans−AM motif assay. Data are presented as mean ± SEM. * indicate a significant difference ( p < 0.05) from Nrf2 +/+ mice.

Article Snippet: Equal concentrations of isolated nuclear protein (30 μg) were employed to assess Nrf2 DNA-binding capability using the TransAM Nrf2 transcription factor assay kit from Active Motif.

Techniques: Gene Expression, Binding Assay, Activity Assay

Journal: Nutrients

Article Title: Accelerated Aging Induced by an Unhealthy High-Fat Diet: Initial Evidence for the Role of Nrf2 Deficiency and Impaired Stress Resilience in Cellular Senescence

doi: 10.3390/nu16070952

Figure Lengend Snippet: Impact of partial Nrf2 ablation and HFD on metabolic parameters. Nrf2 +/+ and Nrf2 +/− mice were fed with standard diet (SD) or high-fat diet (HFD) for 20 weeks. ( A – D ): Final body weight (BW), fasting blood glucose, fasting insulin, and HOMA-IR index measured in Nrf2 +/+ ( n = 7–9) and Nrf2 +/− ( n = 6–7) mice after 20 weeks of respective dietary treatment. Data are presented as mean ± SEM. * denotes a significant difference ( p < 0.05) from the indicated group.

Article Snippet: Equal concentrations of isolated nuclear protein (30 μg) were employed to assess Nrf2 DNA-binding capability using the TransAM Nrf2 transcription factor assay kit from Active Motif.

Techniques:

Journal: Nutrients

Article Title: Accelerated Aging Induced by an Unhealthy High-Fat Diet: Initial Evidence for the Role of Nrf2 Deficiency and Impaired Stress Resilience in Cellular Senescence

doi: 10.3390/nu16070952

Figure Lengend Snippet: Partial Nrf2 ablation increases the expression of senescence markers in peripheral tissues. Analysis of Cdkn2a (p16 INK4a ) gene expression in the liver ( A ) and epididymal white adipose tissue (eWAT) ( B ) samples from Nrf2 +/+ and Nrf2 +/− mice after 20 weeks of consuming a standard diet (SD) or high fat diet (HFD) ( n = 3−6/group). Data are presented as mean ± SEM. * indicates a significant difference ( p < 0.05) from the indicated group.

Article Snippet: Equal concentrations of isolated nuclear protein (30 μg) were employed to assess Nrf2 DNA-binding capability using the TransAM Nrf2 transcription factor assay kit from Active Motif.

Techniques: Expressing, Gene Expression

Journal: Nutrients

Article Title: Accelerated Aging Induced by an Unhealthy High-Fat Diet: Initial Evidence for the Role of Nrf2 Deficiency and Impaired Stress Resilience in Cellular Senescence

doi: 10.3390/nu16070952

Figure Lengend Snippet: Induction of senescence in brain microvascular endothelial cells due to partial Nrf2 ablation. ( A ) Representative images of immunofluorescent labeling used to detect p16-RFP in endomucin+ endothelial cells in the hippocampus of Nrf2 +/+ and Nrf2 +/− mice after 20 weeks of consuming a standard diet (SD) or high fat diet (HFD) ( n = 4/group). White arrows indicate the presence of mRFP positive senescent endothelial cells in the hippocampus. ( B ) Quantification of the p16-RFP+/endomucin+ endothelial cells in the hippocampi of all the groups. Data are presented as mean ± SEM. * p < 0.05.

Article Snippet: Equal concentrations of isolated nuclear protein (30 μg) were employed to assess Nrf2 DNA-binding capability using the TransAM Nrf2 transcription factor assay kit from Active Motif.

Techniques: Labeling

Journal: Nutrients

Article Title: Accelerated Aging Induced by an Unhealthy High-Fat Diet: Initial Evidence for the Role of Nrf2 Deficiency and Impaired Stress Resilience in Cellular Senescence

doi: 10.3390/nu16070952

Figure Lengend Snippet: Schematic illustration of the relationship between consumption of HFD/obesity, oxidative stress, Nrf2 deficiency, and cellular senescence in accelerated aging. Dysfunctional mitochondria, NADPH oxidases, ER stress from misfolded proteins, metabolic processes in peroxisomes, and inflammasomes are major sources of increased reactive oxygen species (ROS) levels in obesity. Nrf2 plays a crucial role in maintaining cellular redox homeostasis. Under conditions of oxidative stress, Nrf2 is released from the Keap1−Cul3−RBX1 complex, after which it translocates into the nucleus, forms heterodimers with small Maf proteins (sMaf), and binds to antioxidant response elements (AREs). This activation leads to the transcription of genes encoding antioxidant enzymes, DNA repair mechanisms and proteins that confer anti-inflammatory effects, facilitating ROS detoxification and restoration of cellular homeostasis. Conversely, impaired Nrf2 activation results in the accumulation of toxic levels of ROS, inducing macromolecular damage like DNA breaks, and triggering senescence, ultimately leading to an accelerated aging phenotype.

Article Snippet: Equal concentrations of isolated nuclear protein (30 μg) were employed to assess Nrf2 DNA-binding capability using the TransAM Nrf2 transcription factor assay kit from Active Motif.

Techniques: Activation Assay