Fig. 4. Effects of the asTGF-ß1 mRNA in culture-activated HSCs. Equal amounts (5 µg/lane) of total cellular RNAs were analyzed by Northern blotting. In the left panel, RNA was isolated from untreated (Lane 1), mock-infected (Lane 2), or Ad5-CMV-asTGF-ß1-infected (Lane 3) rat HSCs. In the right panel, RNA was isolated from mock-infected (Lane 1) or Ad5-CMV-EGFP-infected (Lane 2) cells. RNA was isolated two (2d), three (3d), or four (4d) days after infection. The blots were subsequently hybridized with a 32P-labeled col1(I)-specific probe and the 278-bp PvuII fragment of clone pET3d-rTGF-ß1. Intensities of the 18S and 28S rRNAs and hybridization with a GAPDH-specific cDNA served as controls to verify equivalent loading.