Figure 2

Functional characterization of shRNA-expressing plasmids. (A) HeLa cells were co-transfected with 50 ng of plasmids expressing shRNA targeting firefly luciferase, 200 ng of target pGL2 plasmid and 1 ng of phRL-SV40. Firefly luciferase (FF) activity normalized according to non-targeted Renilla luciferase activity is shown. Firefly luciferase activity in control sample (without a shRNA-expressing plasmid) was set to 1. Values are expressed as mean +/- SEM from samples transfected at least in triplicates. (B) pTMP and pLMP plasmids carrying loxP sites were transformed either to regular or Cre recombinase-expressing E. coli strains. Electrophoresis of isolated plasmid DNA is shown. The recombined plasmid after Cre-mediated recombination is marked by an arrow. (C) HeLa and HEK293 cells were co-transfected with 10-200 ng of plasmids expressing shRNA targeting firefly luciferase, 200 ng of target pGL2 plasmid, and 1 ng of phRL-SV40. Relative firefly luciferase activity compared to control cells is shown. Firefly luciferase activity in the control sample (omitting shRNA-expressing plasmid) was set to 1. Values are expressed as mean +/- SEM from samples transfected at least in triplicates.