Figure 5.

PAI-1 siRNA and shRNAmir suppress the level of PAI-1 protein in normal fibroblasts (N144) transduced with Ad-Wt PAI-1 (100 moi) and reduce collagen accumulation. A: Normal fibroblasts plated at confluent density were transfected with the indicated PAI-1 siRNAs (100 nmol/L) for 24 hours in 10% FBS, then infected with Wt PAI-1 adenoviral vector for 24 hours in 0.1% ITS+, and cultured in 3% FBS for 6 days. Data are presented as the percent expression relative to the effects of the nontargeting control sequence. □, Expression of PAI-1 in the medium presented as the mean ± SD of DNA normalized data (n = 3).

, Accumulation of newly synthesized collagen; n = 3, data handling as in A. B:. Normal fibroblast were infected with a pGIPZ lentiviral vector expressing a shRNAmir targeted to PAI-1, PAI-1–645, or a nontargeting sequence, selected with puromycin, verified by GFP expression, infected with Ad-Wt PAI-1, and cultured as described in A. Nontargeting siRNA: mammalian nontargeting siRNA sequence; PAI-1 siRNAs: siRNA5, siRNA6, or siRNA7. **P ≤ 0.01.