Journal: PeerJ
Article Title: Prokaryotic expression, purification and evaluation of anti-cardiac fibrosis activity of recombinant TGF-β latency associated peptide
doi: 10.7717/peerj.12797
Figure Lengend Snippet: (A) Western blot of the effect of recombinant LAP on α-SMA, Smad2 and p-Smad2 expression in H9C2 cells induced by TGF-β1; +: TGF-β1+LAP group; + 1 : LAP pre-protection group. (B)–(D) Semi-quantitative analysis of α-SMA, Smad2 and p-Smad2 expression presented as the relative ratio to GAPDH. The expression of α-SMA ( P < 0.0001) and p-Smad2 ( P = 0.0003) in the TGF-β1 model group was significantly increased compared with the control group. Compared with the TGF-β1 model group, both TGF-β1+LAP group and LAP pre-protection group significantly reduced the increase of α-SMA (TGF-β1 vs TGF-β1+LAP, P = 0.0409; TGF-β1 vs LAP Pre-protection, P = 0.008) and P-Smad2 (TGF-β1 vs TGF-β1+LAP, P = 0.0285; TGF-β1 vs LAP Pre-protection, P = 0.0044) levels. NC: control group; LAP: 60 μg/mL LAP group; TGF-β1: 10 ng/mL TGF-β1 group; TGF-β1+LAP: Add 10 ng/mL TGF-β1 first and then 60 μg/mL LAP treatment group; LAP pre-protection group: 10ng/mL TGF-β1 and 60μg/mL LAP at the same time. n = 3, *** P < 0.001 vs NC group; # P < 0.05 vs TGF-β1 group; ## P < 0.01 vs TGF-β1 group.
Article Snippet: Then, a rabbit anti-mouse collagen I monoclonal antibody (Affinity Biosciences, Cincinnati, OH, USA, 1:1,000 dilution), a rabbit anti-mouse α-SMA monoclonal antibody (Affinity Biosciences, Cincinnati, OH, USA, 1:1,000 dilution), a rabbit anti-mouse FN monoclonal antibody (Affinity Biosciences, Cincinnati, OH, USA, 1:1,000 dilution), a rabbit anti-mouse Smad2 monoclonal antibody (CST, USA, 1:1,000 dilution) and a rabbit anti-mouse p-Smad2 monoclonal antibody (CST, USA, 1:1,000 dilution) were added separately overnight at 4 °C.
Techniques: Western Blot, Recombinant, Expressing, Control