FIG. 8.

NF-κB activity and transcription of immunoproteasome-related genes and MHC class I genes in THP-1 cells expressing the DV replicon. (A) RT-PCR amplification with 25 cycles using primers for the indicated genes including the β-actin gene as a standard. For each gene the left lane is the amplification product of THP-1 cDNA and the right lane is the amplification product of THP-1.ΔCprME-PAC2A cDNA. Results are from one representative experiment out of two performed. (B) The intensity of the amplification product presented in panel A was quantified and normalized using β-actin. (C) NF-κB activity. A reporter plasmid expressing luciferase through a minimal promoter linked to three copies of the consensus NF-κB-responsive element was used to measure activation of NF-κB. Enzymatic activity in the cell extract was assayed 24 h after transfection. The normalized luciferase activity of THP-1.ΔCprME-PAC2A cells was divided by the normalized luciferase activity of THP-1 and presented relative to NF-κB activity. Results are a summary of two different experiments performed. The bar indicates the standard error (SE). (D) NF-κB activity of K562.ΔCprME-PAC2A cells compared to K562 cells. The experimental procedure is as described for panel C. Results are a summary of two different experiments performed. The bar indicates SE.