Journal: The Journal of Biological Chemistry
Article Title: p75 Neurotrophin Receptor Signaling Activates Sterol Regulatory Element-binding Protein-2 in Hepatocyte Cells via p38 Mitogen-activated Protein Kinase and Caspase-3 *
Figure Lengend Snippet: p75NTR activation induces SREBP2 cleavage in a caspase-3-dependent manner. A, schematic structure of SREBP2 in the membrane, its interaction with the SCAP·INSIG complex, and the protease cleavage sites. In addition to the classical S1P and S2P
Article Snippet: The filters were incubated for 1 h in 50 m m Tris-HCl, pH 7.5, 150 m m NaCl, supplemented with 0.1% Tween 20, and 5% skimmed milk or 5% BSA at room temperature and then overnight at 4 °C with primary antibodies as follows: anti-p75NTR intracellular region (1:1000, Abcam, ab32888); anti-p75NTR extracellular region (1:5000, Millipore, 1554); anti-caspase-3 (1:1000, Cell Signaling, 9665); anti-caspase-3 cleaved form (1:1000, Cell Signaling, 9661); anti-caspase-2 (1:500, Enzo Life Sciences, 11B4); anti-SREBP carboxyl-terminal region (1:500, BD Biosciences, 557037); anti-SREBP amino-terminal region (1:1500, Abcam, ab3062); anti-LDLR (1:2000; Cayman Chemical, 10007665); anti-p38 MAPK (1:1000, Cell Signaling, 9212); anti-phospho-p38 MAPK (1:1000, Cell Signaling, 9216); anti-histone H3 (1:1000, Cell Signaling, 9715); rabbit anti-GFP (1:500, Chemicon); mouse anti-GFP (1:5000, Roche Applied Science); mouse anti-binding immunoglobulin protein (1:1000; BD Biosciences); and anti-β-actin (1:5000; Sigma).
Techniques: Activation Assay