Salamander-Derived, Human-Optimized nAG Protein Suppresses Collagen Synthesis and Increases Collagen Degradation in Primary Human Fibroblasts
Figure 5
Immunofluorescence experiment showing effect of nAG on collagen I (a) and collagen III (b) expression in primary human fibroblasts with or without TGF-β1 treatment. After 48 h of transfection, immunoflourescence was performed by using COL1A1, COL3A1 (primary antibodies), and FITC-conjugated secondary antibody. (a) Immunofluorescence staining of type I collagen is in the following order: control fibroblasts (complete medium + 150 μg/mL L-ascorbic acid), control fibroblasts treated with 3 ng/mL TGF-β1, fibroblasts with nAG plasmid, and fibroblasts with nAG plasmid and treated with 3 ng/mL TGF-β1 (magnification 40x). Note the suppressive effect of nAG on collagen I. (b) Immunofluorescence staining of type III collagen is in the following order: control fibroblasts (complete medium + 150 μg/mL L-ascorbic acid), control fibroblasts treated with 10 ng/mL TGF-β1, fibroblasts with nAG plasmid, and fibroblasts with nAG plasmid and treated with 10 ng/mL TGF-β1 (magnification 40x). Note the complete suppressive effect of nAG on collagen III.
