Time after hypoosmotic stress for Protein kinase B (PKB) phosphorylation to occur

Value 10 min
Organism Human Homo sapiens
Reference Kim RD et al, Activator protein 1 activation following hypoosmotic stress in HepG2 cells is actin cytoskeleton dependent. J Surg Res. 2001 Oct100(2):176-82PubMed ID11592789
Method HepG2 cells were incubated for 1 h with or without 20 microM cytochalasin D, an actin disrupter, and were then exposed for up to 30 min to hypoosmotic medium (200 mOsm/L) to induce swelling. Tumor necrosis factor alpha (1.4 nM) and medium alone served as positive and negative controls, respectively. Western blots measured cytoplasmic phosphorylated or total FAK and PKB. For the detection of PKB, rabbit polyclonal IgGs specific for either total PKB or phosphorylated PKB (Ser473) (New England Biolabs, Beverly MA) were used. Protein extracts from PDGF-treated NIH 3T3 cells served as a control for the phosphorylated PKB (New England Biolabs) antibody binding. All experiments were performed in triplicate.
Comments Researchers hypothesized that hypoosmotic stress-induced activation of PKB and AP-1 in HepG2 cells is dependent on an intact actin cytoskeleton and subsequent FAK phosphorylation
Entered by Uri M
ID 104612