Figure 4

K13 activates HIV LTR through the classical NF-κB pathway. A. 293T cells were transfected with an empty vector or K13 along with an HIV LTR/luciferase reporter construct and a β-galactosidase reporter construct as described in Fig. 1A. The amount of IκBαSS32/36AA inhibitor plasmid (500 ng/well) was five times the amount of vector (pcDNA3) or K13 (100 ng/well) plasmid. The values shown are averages (Mean ± S.E.) of one representative experiment out of three in which each transfection was performed in duplicate. B. Western blot analysis showing siRNA-mediated knock-down of p65, c-Rel and RelB expression. The blot was re-probed with a monoclonal antibody against actin (bottom panel) to show equal loading of all lanes and specificity of gene silencing. C. 293T cells were transfected with an empty vector or a K13 expression plasmid along with a control siRNA oligo-duplexes or siRNA duplexes against c-Rel, p65 and RelB, respectively. The luciferase reporter assay was performed as described in Fig 1A.