Fig. 1.

Vectors used in the construction of recombinant plasmids. (A) The Lentiviral pCDH vector contains common domains such as selection antibiotics, copGFP expression cassette under the control of the EF1βα promoter, and the cytomegalovirus (CMV) promoter that leads to the production of our selected viral miRNAs inserted between the EcoRI and BamHI sites. (B) psiCHECK-2 vector designed to provide quantitative and rapid optimization of RNA interference. Renilla luciferase was used as the primary reporter gene, and the 3’-UTR of SMAD3 and SMAD4 was separately cloned into the XhoI and NotI sites located downstream of the Renilla translational stop codon. Introduction of firefly luciferase in this vector, as a second reporter gene, allows the normaliztion of Renilla luciferase expression, achieving robust and reproducible results