Journal: Journal of Biological Chemistry

Article Title: α-Thrombin Rapidly Induces Tyrosine Phosphorylation of a Novel, 74–78-kDa Stress Response Protein(s) in Lung Fibroblast Cells

doi: 10.1074/jbc.m409043200

Figure Lengend Snippet: FIG. 4. The p74–78-kDa protein is localized to the cytoplasm. A, serum-starved cells were untreated or treated -thrombin (0.4 units/ml; 15 min) and lysed, and then membrane, cytoplasm, and nuclear frac- tions were prepared as described under “Experimental Procedures.” 15 g of the sample were run on an 8% SDS-polyacrylamide gel, trans- ferred to nitrocellulose, and immunoblotted with the phospho-specific anti-Stat3 antibody. The positions of three distinct bands (74–78 kDa) are indicated by arrows. B and C are blots probed with anti-Stat3 antibody and anti-TGF- receptor antibody, respectively. These blots are representative of three independent experiments. Th, -thrombin; CR, cross-reactive.

Article Snippet: IL-6, PDGF, transforming growth factor- (TGF- ), and basic fibroblast growth factor (FGF) were obtained from R & D Systems; nitrocellulose membrane was from Amersham Biosciences; phospho-specific antiStat3 antibody was from BIOSOURCE; anti-Stat3 antibody and antiE2F-1 antibody were from Santa Cruz Biotechnology; anti-phosphotyrosine antibody was from Upstate Biotechnology, Inc.; anti-GRP antibody was from Oxford Biomedical Research; goat anti-rabbit IgG and rabbit anti-mouse IgG were from Bio-Rad; all other chemicals were either from Sigma or Fisher.

Techniques: Membrane