Skip to main content
Figure 5 | Retrovirology

Figure 5

From: Kaposi's sarcoma associated herpes virus-encoded viral FLICE inhibitory protein activates transcription from HIV-1 Long Terminal Repeat via the classical NF-κB pathway and functionally cooperates with Tat

Figure 5

Mechanism of K13-induced HIV-1 LTR activation. A. Role of IKK complex in K13-induced HIV-1 LTR reporter activity. Wild-type and IKK-deficient cells were transiently transfected with an empty vector or K13 expression plasmid (500 ng/well) along with an HIV/luciferase reporter construct (100 ng/well) and a synthetic Renilla luciferase (phRL-TK) reporter vector (75 ng/well) by using LIPOFECTAMINE 2000 Reagent (Invitrogen, Carlsbad, CA) according to manufacturer's instruction. Thirty-six hours after transfection, cell lysates were used for reporter assays. Luciferase activity was normalized relative to the Renilla luciferase activity to control for the difference in the transfection efficiency. The values shown are averages (Mean ± S.E.) of one representative experiment out of three in which each transfection was performed in duplicate. B. Inhibitors of the NF-κB pathway significantly block K13-actived HIV LTR promoter. 293T cells were transfected with the empty vector or K13 along with an HIV LTR/luciferase reporter construct and a β-galactosidase reporter construct as described in Fig.1A. Eight hours after transfection, cells were treated with DMSO or different inhibitors for 24 h and then lysed for the reporter assay. The values shown are averages (Mean ± S.E.) of one representative experiment out of three in which each transfection was performed in duplicate. C. Effect of Murr1 on K13-induced HIV-1 LTR activation. 293T cells were transfected with an empty vector (pCDNA3) or K13 along with an HIV LTR/luciferase reporter construct and a β-galactosidase reporter construct as described in Fig. 1A. The amount of Murr1 plasmid (500ng/well) was five times the amount of vector or K13 (100ng/well). The values shown are averages (Mean ± S.E.) of one representative experiment out of three in which each transfection was performed in duplicate.

Back to article page