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. 2012 Oct 23;21(1):8–17. doi: 10.1038/mt.2012.135

Figure 1.

Figure 1

Immunoblot analysis and bioactivity assays for TGFβ3 produced from recombinant adenoviral constructs. (a) Conditioned media were collected from COS7 cell cultures transduced with an empty adenoviral vector (Ad-Null) and the vectors encoding for latent TGFβ3 (Ad-TGFβ3), GFP (Ad-GFP) or GFP, and latent TGFβ3 (Ad-GFP-TGFβ3), and analyzed on an immunoblot with a TGFβ3-specific antibody. The last lane is loaded with fully processed 25 kDa recombinant human TGFβ3 (rhTGFβ3), as indicated with an arrowhead. (b) Bioactivity of virally expressed TGFβ3 was assayed (N = 3), using mink lung epithelial (MLE) cells stably transfected with a luciferase reporter construct under the control of a TGFβ-responsive plasminogen activator inhibitor-1 promoter. Conditioned media collected from COS7 cell cultures, which had been infected with Ad-GFP-TGFβ3, Ad-GFP, Ad-TGFβ3, or Ad-Null, were assayed for luciferase reporter activity in MLE cells. Bovine serum albumin (BSA) and rhTGFβ3 (10 and 100 nmol/l) were used in control assays. GFP, green fluorescent protein; TGFβ3, transforming growth factor β3.