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GeneWorks
sequence-centered analysis software Sequence Centered Analysis Software, supplied by GeneWorks, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more https://www.bioz.com/result/sequence-centered analysis software/product/GeneWorks Average 90 stars, based on 1 article reviews
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Novus Biologicals
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Image Search Results
Journal: Nature Communications
Article Title: Hydralazine induces stress resistance and extends C. elegans lifespan by activating the NRF2/SKN-1 signalling pathway
doi: 10.1038/s41467-017-02394-3
Figure Lengend Snippet: SILAC screen identifies NRF2 pathway activation with hydralazine treatment. a The SILAC workflow for identification of pathways activated by hydralazine ( n = 2). b Proteins with their log2 ratios (treated/untreated) were mapped into the ingenuity pathway analysis (IPA) database where the NRF2 pathway was found activated. Raw MS data for proteins SQSTM1, FTH1, and GSTK1 are also shown as part of the manual validation of the data. c NRF2 pathway activation reported by IPA signified with Z score of 0.156 and p value of 1.97E−10, right-tailed Fisher exact test
Article Snippet: After blocking the membrane with 5% milk for 1 h at room temperature, the membrane was incubated further for 2 h with antibodies specific for target proteins:
Techniques: Multiplex sample analysis, Activation Assay, Biomarker Discovery
Journal: Nature Communications
Article Title: Hydralazine induces stress resistance and extends C. elegans lifespan by activating the NRF2/SKN-1 signalling pathway
doi: 10.1038/s41467-017-02394-3
Figure Lengend Snippet: Hydralazine enhances NRF2 signaling in SH-SY5Y cells. a Hydralazine increased cellular NRF2 protein in a dose-dependent manner demonstrated by western blot analysis of cell lysates (input) and NRF2 immunoprecipitates (IP:NRF2). * p < 0.05 and ** p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. b The mass spectrometry based label-free quantification of NRF2 immunoprecipitates prepared in part A. Extracted ion chromatogram of a NRF2 peptide, IINLPVVDFNEM ox M ox SK, as well as bar plot quantification are shown. Samples were treated with 0.05% H 2 O 2 prior to mass spectrometric analysis. * p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. c Hydralazine reduced the interaction between NRF2 and KEAP1. Interactions were measured by reciprocal Co-IPs followed by western blot analysis. * p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. d NRF2 translocates to the nucleus with hydralazine treatment. Treated cells were subjected to cell fractionation and western blot analysis. * p < 0.05 and ** p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. e Hydralazine treatment increased nuclear NRF2 phosphorylation quantified using an antibody specific to NRF2 phosphorylation at serine 40. * p < 0.05 and ** p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. f TXN, a potent regulator of the NRF2–KEAP1 response system, is upregulated with hydralazine treatment. * p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. g ARE-driven luciferase activity was increased with hydralazine treatment, indicating an increase in the transcriptional activation of NRF2 target genes. Hydrazine did not increase luciferase activity. ** p < 0.01, two-tailed Student's t test, n = 6, mean ± SD. h Hydralazine treatment increased the expression of NRF2 downstream targets measured by qPCR using actin as internal control. For the list of primers used for qPCR, see Supplementary Table online. * p < 0.05, and ** p < 0.01, two-tailed Student's t test, n = 6, mean ± SEM. i Hydralazine treatment increased the expression of NRF2 downstream target measured by western blot analysis. * p < 0.05, two-tailed Student's t test, n = 3, mean ± SD
Article Snippet: After blocking the membrane with 5% milk for 1 h at room temperature, the membrane was incubated further for 2 h with antibodies specific for target proteins:
Techniques: Western Blot, Two Tailed Test, Mass Spectrometry, Quantitative Proteomics, Cell Fractionation, Phospho-proteomics, Luciferase, Activity Assay, Activation Assay, Expressing, Control
Journal: Nature Communications
Article Title: Hydralazine induces stress resistance and extends C. elegans lifespan by activating the NRF2/SKN-1 signalling pathway
doi: 10.1038/s41467-017-02394-3
Figure Lengend Snippet: SKN-1 pathway is activated with hydralazine treatment in C . elegans . In all experiments animals were treated with 100 µM hydralazine for 72 h unless otherwise stated. a Fluorescent photomicrograph showing hydralazine treatment increases SKN-1::GFP localization in the intestinal nuclei in geIs10 transgenic animals. Scale bar = 50 µm. b Quantification of SKN-1 intestinal nuclear accumulation represented as percentage of worms with high (≥15 GFP-positive intestinal nuclei), medium (5–15 GFP-positive intestinal nuclei), or low (≤5 GFP-positive intestinal nuclei) nuclear SKN-1::GFP. ** p < 0.01, two-tailed Student's t test, n = 100 three independent trials. c Fluorescent photomicrograph showing GFP signal of SKN-1B in ASI neurons of ldIs7 transgenic worms. GFP signal intensity in ASI neurons increased with hydralazine treatment as quantified with Image J software. Scale bar = 20 µm. ** p < 0.01, two-tailed Student's t test, n = 75 four independent trials, mean ± SD. d GFP signal intensity quantification showing upregulation of GST-4::GFP in the intestine of hydralazine-treated dvIs19 transgenic worms after 48 h. ** p < 0.01, two-tailed Student's t test, n = 35 three independent trials, mean ± SD. e Fluorescent photomicrograph showing hydralazine treatment did not increase GST-4::GFP in skn-1 RNAi fed dvIs19 transgenic animals. Scale bar = 80 µm. ** p < 0.01, two-tailed Student's t test, n = 30 three independent trials, mean ± SD. f The level of superoxide (O 2 •− ) measured with DHE fluorophore signal intensity decreased in wild-type C. elegans treated with hydralazine but not in SKN-1 mutant worms. * p < 0.05, two-tailed Student's t test, n = 30 three independent trials, mean ± SD. g A volcano plot showing activation of the SKN-1/NRF2 pathway in wild-type C. elegans using Tukey’s honestly significant difference test. Proteins were quantified in both treated and untreated animals using label-free mass spectrometry and ratios were uploaded for identification of activated pathways via IPA analysis. The SKN-1/NRF2 pathway was among the activated pathways with Z score of 3.317 and p value of 0.0002, right-tailed Fisher exact test. A list of human orthologs of SKN-1 pathway members and their Log FC is shown in a table below the plot
Article Snippet: After blocking the membrane with 5% milk for 1 h at room temperature, the membrane was incubated further for 2 h with antibodies specific for target proteins:
Techniques: Transgenic Assay, Two Tailed Test, Software, Mutagenesis, Activation Assay, Mass Spectrometry