Sirna Constructs Targeting P22 Phox, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Glucose deprivation activates a metabolic and signaling amplification loop leading to cell death"
Article Title: Glucose deprivation activates a metabolic and signaling amplification loop leading to cell death
Journal: Molecular Systems Biology
Figure Legend Snippet: NADPH oxidase- and mitochondria-derived ROS contribute to glucose withdrawal-induced phospho-tyrosine signaling. ( A ) Inhibition of NOX inhibits glucose withdrawal-induced signaling. LN18, T98, and U87 cells were starved of glucose and pyruvate in the presence of DMSO or DPI (1 μM). Western blotting demonstrated that NOX activity is required for the induction of phospho-tyrosine signaling. ( B ) Knockdown of the NOX subunit p22 phox attenuates phospho-tyrosine signaling following glucose withdrawal. U87 cells were reverse transfected with control, non-targeting siRNA or siRNA against p22 phox , DUOX1 or DUOX2. Forty-eight hours later, cells were starved of glucose and pyruvate for 5 h. Western blotting demonstrated that knockdown of p22 phox but not DUOX1/2 attenuated glucose withdrawal-induced phospho-tyrosine signaling. p22 phox knockdown efficiency was >90% . ( C ) LN18, T98, and U87 cells were starved of glucose and pyruvate in the presence of either DMSO or BAPTA-AM (25 μM). Western blotting with an anti-phospho-tyrosine antibody demonstrated that chelation of intracellular Ca 2+ by BAPTA-AM completely abrogated glucose withdrawal-induced phospho-tyrosine signaling. Treatment with extracellular EDTA (25 μM) had no effect. Actin and GRB2 served as equal loading controls. ( D ) ρ 0 derivatives of T98 and U87 cells do not exhibit upregulation of phospho-tyrosine signaling or activation of Src in response to glucose withdrawal. ( E ) Catalase rescues parental but not ρ 0 cells from glucose withdrawal-induced cell death. Cells were starved of glucose and pyruvate with or without catalase (1 kU/ml), and viability was measured by Trypan blue exclusion 24 h later.
Techniques Used: Derivative Assay, Inhibition, Western Blot, Activity Assay, Transfection, Activation Assay