Fig 6.

Knockdown of β-catenin and TCF-4 modulates C/EBP and NF-κB tethering on HIV LTR. PDAs at 60 to 70% confluence were transfected with β-catenin siRNA, TCF-4 siRNA, or a scrambled siRNA negative control (NC). After 48 h, cells were transiently transfected with an LTR-Luc construct. At 24 h post-LTR transfection, cells were cross-linked with 1.0% formaldehyde and sonicated to generate chromatin preparations (5 × 10⁶ cells/sample). ChIP was performed using antibodies against C/EBPβ, C/EBPδ, or NF-κB p65 subunit (5 μg antibody/IP). Data shown are normalized to a nontargeting IgG control. *, P < 0.05 in comparison with control siRNA. (d) PDAs at 60 to 70% confluence were transfected with β-catenin siRNA, TCF-4 siRNA, or a scrambled siRNA negative control (NC). After 48 h, cells were transfected with an NF-κB luciferase reporter plasmid, and a luciferase assay was performed after an additional 24 h. Results are reported as fold change in relative light units (RLU) compared to the negative control and are normalized to the amount (micrograms) of cellular protein. All data represent at least three independent experiments performed in duplicate. *, P < 0.05 in comparison with the negative control.