Figure 4

Lentiviral siRNA silencing of NFκB reduces RNA readthrough of LV SIN LTR. The NF kB siRNA lentiviral vector was made by annealing two oligos pre-treated at 95°C for 5 min and gradually cooled down to room temperature. The annealed products were cloned into pTYF-Puro-hU6r LV-driven by a human U6 promoter. The NFκB silenced TE26 cell line was generated after puromycin selection and used for RNA TR analysis as described in the article. A. Real-time quantitative RT-PCR analysis of NFκB mRNA. B. Western analysis of NFκB expression in LV-NFκB siRNA-transduced TE26 cells. C. Comparison of RNA readthrough rate in TE26 and NFκB-silenced TE26 cells. The significance of difference is analyzed by paired student t test and shown by asterisks.