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. 2010 Oct 8;5(10):e13172. doi: 10.1371/journal.pone.0013172

Figure 3. Reconfirmation of the inhibitory potential of selected shRNA-sequences.

Figure 3

a) ShRNAs were PCR-amplified from genomic DNA of selected cell clones as shRNA expression cassettes consisting of H1 promoter, shRNA-sequence and polyT. Each of the 200 individual cassettes were co-transfected with the HIV-1 specific Luciferase reporter construct pNL4.3Luc.R-E- into HEK 293 FT cells. Luciferase expression was measured 48 h p.t. Cells transfected with pNL4.3Luc.R-E- and a scrambled shRNA (sh scr) expressing cassette were used as control. b) Map of the identified shRNAs and re-evaluation of their inhibitory potential upon co-transfection as described in a). Cells which were transfected with pNL4.3LucR-E- alone or co-transfected with a scrambled siRNA control served as controls. Error bars indicate +/− SD of mean of three independent experiments.