Journal: Experimental Biology and Medicine
Article Title: Inhibition of Mastermind-like 1 alleviates liver fibrosis induced by carbon tetrachloride in rats
doi: 10.1177/1535370218810892
Figure Lengend Snippet: Effect of MAML1-siRNA on Notch and Wnt/β-catenin signal transduction in HSCs. HSC-T6 cells were treated with MAML1-siRNA or control siRNA. Then, the expression of Hes1 (a), Cyclin D1 (b) and c-Myc (c) was detected using real-time PCR and Western blot. Additionally, HSC-T6 cells were cultured in 96-well plates. Lipofectamine 2000 was used to co-transfect with 60 ng of reporter vector (TCF/LEF (d) or CSL (e) luciferase and Renilla luciferase) or negative control reporter and 20 nM of MAML1-siRNA or negative control siRNA; 48 h after transfection, luciferase activities were measured by the Dual Luciferase Assay System. Data represent the results of three independent experiments. *P < 0.05 versus the normal HSC-T6 or control siRNA group.
Article Snippet: Luciferase reporter assay Wnt/β-catenin pathway (TCF/LEF) reporter kit, Notch pathway (CSL) reporter kit and Dual Luciferase (Firefly-Renilla) Assay System (BPS Bioscience) were used to monitor the activity of Wnt/β-catenin or Notch signal in HSC-T6 cells following the protocol provided by BPS Bioscience.
Techniques: Transduction, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Plasmid Preparation, Luciferase, Negative Control, Transfection