Journal: Oxidative Medicine and Cellular Longevity

Article Title: Attenuation of β -Amyloid-Induced Oxidative Cell Death by Sulforaphane via Activation of NF-E2-Related Factor 2

doi: 10.1155/2013/313510

Figure Lengend Snippet: SUL-induced activation of Nrf2 in SH-SY5Y cells. Cells were incubated with SUL (5 μ M) for indicated times and nuclear as well as total protein sample extracts were prepared. (a) Nuclear translocation of Nrf2 was monitored by western blotting of nuclear extracts by probing with anti-Nrf2 specific antibody. (b) Nrf2-ARE binding activity was measured by EMSA according to the manufacturer's instruction using biotin-labeled oligonucleotide specific for Nrf2. (c) Phosphorylation of Nrf2 was assessed by western blot analysis with anti-phospho-Nrf2 (Ser40) antibody. The levels of Topo II (a) and actin (c) were examined to ensure equal amount of nuclear and total protein as loading controls, respectively.

Article Snippet: Subsequently, biotin-labeled oligonucleotide specific to Nrf2 (5′-TGGGGAACCTGTGCTGAGTCACTGGAG-3′, Panomics, CA, USA) was added to the reaction mixture and additionally incubated for 10 min at RT.

Techniques: Activation Assay, Incubation, Translocation Assay, Western Blot, Binding Assay, Activity Assay, Labeling, Phospho-proteomics

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Attenuation of β -Amyloid-Induced Oxidative Cell Death by Sulforaphane via Activation of NF-E2-Related Factor 2

doi: 10.1155/2013/313510

Figure Lengend Snippet: Effect of Nrf2 gene knock-down on SUL-mediated protection against A β 25–35 -induced apoptotic cell death. SH-SY5Y cells were transiently transfected with siRNA of Nrf2 according to the protocol provided by manufacturer and then exposed to A β 25–35 (15 μ M) in the presence or absence of SUL (5 μ M) for 24 h. (a) MTT assay was performed to measure cell viability. Data are represented as mean ± S.D. ( n = 3). ** P < 0.01, significantly different between groups. (b) TUNEL staining was conducted to verify DNA fragmentation in situ . (A) Vehicle-treated control; (B) A β 25–35 alone (15 μ M); (C) A β 25–35 (15 μ M) + SUL (5 μ M); (D) A β 25–35 (15 μ M) + SUL (5 μ M) + Nrf2-siRNA. (c) TMRE staining was performed to compare MMP. Data are represented as mean ± S.D. ( n = 3). ** P < 0.01, significantly different between groups. (d) Protein expression of Bcl-2 and Bax was determined by western blot analysis using specific antibodies. Actin levels were examined to ensure equal amount of protein loading.

Article Snippet: Subsequently, biotin-labeled oligonucleotide specific to Nrf2 (5′-TGGGGAACCTGTGCTGAGTCACTGGAG-3′, Panomics, CA, USA) was added to the reaction mixture and additionally incubated for 10 min at RT.

Techniques: Knockdown, Transfection, MTT Assay, TUNEL Assay, Staining, In Situ, Control, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Attenuation of β -Amyloid-Induced Oxidative Cell Death by Sulforaphane via Activation of NF-E2-Related Factor 2

doi: 10.1155/2013/313510

Figure Lengend Snippet: Schematic diagram that describes neuroprotective effects of SUL against A β -induced oxidative cell death in AD. SUL attenuates A β -induced oxidative damages, pro-apoptotic signals, and apoptotic cell death through the activation of Nrf2-ARE signaling pathway, which consequently fortify Nrf2-dependent antioxidant defense capacity.

Article Snippet: Subsequently, biotin-labeled oligonucleotide specific to Nrf2 (5′-TGGGGAACCTGTGCTGAGTCACTGGAG-3′, Panomics, CA, USA) was added to the reaction mixture and additionally incubated for 10 min at RT.

Techniques: Activation Assay