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. 2009 Mar 16;4(3):e4875. doi: 10.1371/journal.pone.0004875

Figure 3. Casp8p43 induces NF-κB dependent HIV LTR transcription.

Figure 3

(A) Jurkat T cells were treated with gp120 or CH11 anti-Fas antibody and analyzed for caspase 8 cleavage by western blot. (B) Jurkat T cells transfected with κB luciferase reporter plasmid, TK-Renilla as a control for transfection efficiency, and either control vector or vector expressing full length caspase 8, or p43 subunit of caspase 8. As a positive control, NF-κB elements p65/p50 were used as indicated. Results are representative of three independent experiments. (C) Jurkat T cells were transfected with HIV LTR or HIV LTR missing the KB motifs (HIV LTR Δ KB), along with TK-Renilla as a transfection control, and the Casp8p43 with or without IKKγ, dominant negative (DN) IκBα as indicated. Data are expressed as Luciferase units, normalized to TK-Renilla and representative of three independent experiments. (D) 293T cells were transfected with the indicated constructs. The following morning nuclear extracts were prepared and incubated with a P32 labeled HIV LTR NF-κB probe in the presence or absence of antibodies to NF-κB proteins p50 and p65. Then complexes were run on a 6% nondenaturing gel, and analyzed using autoradiography.