Overexpression and knockdown of HIF-lα in NB cells by lentiviral vectors.

Abstract

<p>(A, D) SH-SY5Y and IMR32 cells were transfected with lentiviral vectors and observed by fluorescence microscopy after 72 h. More than 90% of the cells expressed green fluorescent protein (GFP). Magnification, ×200. (B, E) Western blot analysis showed significantly higher HIF-lα expression in SH-SY5Y and IMR32 cells transfected by Lenti-HIF-lα than in cells transfected by Lenti-GFP after 72 h under normoxia. β-actin was used as a loading control. (C, F) SH-SY5Y and IMR32 cells were transfected with Lenti-scrambled siRNA and three different siRNAs targeting HIF-lα. After a 72-h-transduction, an additional 8-h-hypoxic treatment was needed before Western blotting. siRNA2 was proven to be the most effective one of the three siRNAs. β-actin was used as a loading control. N, normoxia; H, hypoxia.</p