Lentiviral-Mediated RNA Interference against TGF-Beta Receptor Type II in Renal Epithelial and Fibroblast Cell Populations In Vitro Demonstrates Regulated Renal Fibrogenesis That Is More
Efficient than a Nonlentiviral Vector
Figure 1
Gene silencing of EGFP by transfection of lentiviral and non-lentiviral expression constructs in vitro. RNAi EGFP gene silencing was tested at 24, 48, and 72 hrs in EGFP stably expressing S7 cells post transfection of an EGFP-RNAi expression construct (pPlasRi-EGFP) and a lentiviral RNAi construct (pLentiRi-EGFP). Fluorescence microscope images of cultured cells are demonstrated as well as the FACS analysis. The percent of EGFP positive (+) cells (mean ± SEM) was assessed using FACS results, and these are shown graphically. There was a significant decrease of EGFP 24 hrs after both transfections compared to controls (** in comparison with controls). The pPlasRi-EGFP plasmid showed a transient silencing effect in comparison with a persistent inhibition by the pLentiRi-EGFP, which produced a continuing inhibition of EGFP (+) cells at 48 and 72 hrs. Gene silencing of EGFP by the pLentiRi-EGFP plasmid remained at 95.91% at 72 hrs.
