A parapoxviral virion protein targets the retinoblastoma protein to inhibit NF-κB signaling - Fig 4

<p>(A) Effect of ORFV119 on NF-κB mediated gene expression. OFTu cells were infected with OV-IA82, OV-IA82-Δ119, or OV-IA82-RV119^Flag (MOI, 10), and levels of TNFα, TLR2, NF-κB1 and IL36α mRNA assessed by real-time PCR at 2 h p.i. Fold changes are relative to OV-IA82 treatment. Results are the mean values of three independent experiments. (*, <i>P<0</i>.<i>05</i>; **, <i>P<0</i>.<i>01</i>). (B)—Effect of ORFV119 on NF-κB-p65 phosphorylation and nuclear translocation. OFTu cells were mock infected or infected with OV-IA82, OV-IA82-Δ119, OV-IA82-RV119^Flag, or OV-IA82-RV119^{Lx<u>G</u>xE-Flag} (MOI, 10), fixed at 1 h p.i, sequentially incubated with primary antibody against NF-κB-p65 and appropriate Alexa Fluor 488 or 594 labeled secondary antibodies, counterstained with DAPI, and examined by confocal microscopy. Green/Red, NF-κB-p65; Blue, DAPI. Results are representative of three independent experiments. (C) OFTu cells were infected and processed as in B, and the percentage of cells exhibiting nuclear NF-κB-p65 calculated at the indicated time points p.i. Results are shown as mean ± S.D. (n = 300 cells/slide) for three independent experiments [*, <i>P<0</i>.<i>05</i> (OV-IA82 vs OV-IA82-Δ119 and OV-IA82 vs OV-IA82-RV119^{Lx<u>G</u>xE-Flag}), ^#<i>P<0</i>.<i>05</i>, ^##<i>P<0</i>.<i>01</i> (OV-IA82-Δ119 Vs OV-IA82-RV119^{Lx<u>G</u>xE-Flag})].