Fig 1.

Recombinant elastin expression in rat BMSCs. (A) Cultured rat BMSCs were effectively transduced with the adenovector, as indicated by GFP fluorescence in Ad-CMV-GFP transduced cells at day 4 (40× magnification). (B) Human elastin mRNA in the BMSCs was evaluated by RT-PCR 2 and 4 days after transduction, with primer sets specific for human and rat elastin. (C) Western blotting with an antibody specific for human tropoelastin confirmed the expression of elastin protein in the Ad-CMV-elastin transduced rat BMSCs. (D) Immunostaining with DAPI, anti-human elastin antibody and FITC-conjugated phalloidin (for F-actin) revealed no detectable human elastin in the culture of BMSCs transduced with empty vector (left panels). However, BMSCs transduced with Ad-CMV-elastin (right panels) showed a network of human elastin molecules (200× magnification). As the strong elastin staining in the Ad-CMV-elastin group overshadowed the F-actin staining, we enhanced the contrast/brightness of the F-actin staining to show that most of the elastin-staining cells were F-actin positive. (Con = empty vector; GFP = Ad-CMV-GFP; representative of three experiments).