Salamander-Derived, Human-Optimized nAG Protein Suppresses Collagen Synthesis and Increases Collagen Degradation in Primary Human Fibroblasts
Figure 2
Western blot assay (a) and immunofluorescence (b) experiments showing nAG protein expression in primary human fibroblasts. (a) Western blot was performed after 24 h and 48 h of transfection. The cells were lysed by using RIPA cocktail; proteins were separated on 12% SDS polyacrylamide gel. The primary antibody V5 probe and HRP-conjugated secondary antibody were used for nAG protein detection in the following lanes: nontransfected fibroblasts (negative control 1), fibroblasts with nAG plasmid without lipofection (negative control 2), nAG transfected fibroblasts tested after 24 h and nAG transfected fibroblasts tested after 48 h. (b) Immunofluorescence assay was performed after 48 h of transfection and cells were fixed and permeabilized by 2% PFA/0.1% Triton x-100. V5 probe was used as primary antibody and green fluorescence was detected by using FITC-conjugated secondary antibody. Compare nAG protein expression in nontransfected fibroblasts (negative control) to nAG expression in transfected fibroblasts (magnification 40x).
