Fig. 3. Cell proliferation and quantitative determination of -SMA, LTBP-1, TGF-ß1, TGF-ß2, TGF-ß3, TßRI, and TßRII. A, proliferation of mock-, Ad5-CMV-asTGF-ß1-, or Ad5-CMV-EGFP-infected HSCs. HSCs were seeded in DMEM containing 10% FCS, and at the second day of culture, the serum was reduced to 5% heat-inactivated FCS and infected with indicated adenoviruses. Twenty-four h later, the medium was changed to 0.5% FCS for 24 h, and then [3H]thymidine (75 kBq/ml) was added. Additionally, the cells were stimulated with human recombinant PDGF-BB or left unstimulated. The mean values of triplicate determinations of a representative experiment are shown; bars, SD. B, quantitative determination by cell ELISA of -SMA and LTBP-1. C, immunoblot detection of -SMA protein levels in untreated HSC (Lane 1), mock-infected (Lane 2) or Ad5-CMV-asTGF-ß1-infected (Lane 3) HSCs. Protein extracts were isolated three (3d) or five (5d) days after infection. D, immunoblot detection of p38-MAPK in untreated (Lane 1), mock-infected (Lane 2) or Ad5-CMV-asTGF-ß1-infected (Lane 3) HSCs. E and F, quantitative analysis of TGF-ß isoforms (E) and type I and type II receptors (F) as determined by cell ELISA. The concentrations of individual proteins are given as fluorescence units (FU) and refer to the DNA content of control cultures.