Western blot analysis is shown using Collagen type I antibody to detect expression of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARgamma-transduced cell lysates (right lane). Protein staining shown below each blot depicts equal protein loading. An equal amount of the whole cell protein (100ug) was separated by SDS-PAGE and electroblotted to nitrocellulose membranes. Proteins were detected by incubating the membrane with Collagen type I antibody at a concentration of 0.2-2ug/10 ml in TBS (100 mM Tris-HCl, 0.15 M NaCl, pH 7.4) with 5% Non-fat milk. Detection occurred by incubation with a horseradish peroxidase-conjugated secondary antibody at 1ug/10 ml. Proteins were detected by a chemiluminescent method using the PIERCE ECL kit (Amersham Biosciences). Other detection systems will yield similar results. See Hazra et al. (2004) for additional details.
