Figure 7

Imp7 KD inhibits plasmid DNA transfection. (A) HeLa DxR KD (shDxR), imp7 KD (shimp7) and imp7 back complemented cells (shimp7+7R) were plated onto 6-well plates and transfected the next day with different amounts of the linearized HIV-1 plasmid DNA (pHR') complexed with FuGene 6. Twenty-four hours after transfection, the percentage of GFP+ cells was measured by flow cytometry. (B) HeLa DxR KD cells (DxR) and two different imp7 KD clones (please refer to Figure 2D) were plated onto 24-well plates and transfected as described in panel A. GFP+ cells were counted twenty four hours after transfection by flow cytometry. Note that 1/4 of the DNA was used in experiments shown in panel B compared to panel A because cells were plated onto 24-well plates. Bars represent the average value ± SD of three independent experiments. (C) HeLa DxR and imp7 KD cells were plated into 6 well plates with or without 1.5 micrograms/ml aphidicolin for 24 hours. Cells were then transfected with the indicated amount of linearized plasmid DNA (pHR') as described in (A) and analyzed by flow cytometry 24 hours later. Data are expressed as fold inhibition relative to shDxR cells and are representative of two independent experiments.