Figure 5

Effect of hStau-2 over-expression and knockdown on Rev export activity. A) Over-expression of hStau-2 increased RRE-RNA levels in the cytoplasmic fraction of HEK293T cells: hStau-2 over-expressed HEK293T cells or control cells were transfected with pNL4-3 pro-viral DNA. After 24 hrs of pNL4-3 transfection, total RNA was isolated from the cytoplasmic fractions and cDNA was prepared from 500 ng of RNA. Viral RRE was measured in the cytoplasmic fraction by absolute qRT-PCR. B) A leftward shift in CT curves for RRE-containing RNA upon hStau-2 overexpression. C) hStau-2 knockdown reduced RRE-RNA levels in the cytoplasmic fraction of HEK293T cells: hStau-2 siRNA or scrambled siRNA were transfected into HEK293T cells followed by pNL4-3 pro-viral DNA transfection. After 24 hrs of pNL4-3 transfection, total RNA was isolated from the cytoplasmic fractions and cDNA was prepared from 500 ng of RNA. Viral RRE was measured in the cytoplasmic fraction by absolute qRT-PCR. D) A rightward shift in CT curves for RRE-containing RNA upon hStau-2 knockdown. The experiments were done more than 3 times and the error bar represents ± SD. *p value ≤ 0.05 was taken as significant.