Salamander-Derived, Human-Optimized nAG Protein Suppresses Collagen Synthesis and Increases Collagen Degradation in Primary Human Fibroblasts
Figure 4
Western blot experiment for detection of collagen I (a) and collagen III (b) expression in primary human fibroblasts transfected with nAG plasmid with or without treatment with TGF-β1. Cells were lysed after 48 hours using RIPA cocktail. Proteins were separated on 7.5% SDS polyacrylamide gel and detected by using COL1A1 (primary antibody for collagen I), COL3A1 (primary antibody for collagen III), and HRP-conjugated secondary antibody and the immunoblots were visualized by using ECL kit. (a) Collagen I detection: lane-1: collagen I expression in non-transfected fibroblasts without any treatment (control 1), lane-2: collagen I expression in fibroblasts transfected with nAG plasmid, lane-3: collagen I expression in non-transfected fibroblasts treated with TGF-β1 (control 2), lane-4: collagen I expression in fibroblasts transfected with nAG plasmid and treated with TGF-β1, and lanes-5–8: internal control β-actin. (b) Collagen III detection: lane-1: collagen III expression in non-transfected fibroblasts without any treatment (control 1), lane-2: collagen III expression in non-transfected fibroblasts with TGF-β1 treatment (control 2), lane-3: collagen III expression in fibroblasts transfected with nAG plasmid, lane-4: collagen III expression in fibroblasts transfected with nAG plasmid and treated with TGF-β1, and lanes-5 to 8 are internal control β-actin.
