Extended Data Fig. 6: F14 is unique among known viral inhibitors of NF-κB.
(a) Top: Amino acid sequence of the TAD of HSV-1 VP16 with the acidic activation domain similar to p65 highlighted in red, and hydrophobic residues (Φ) are indicated. Middle: NF-κB-dependent luciferase activity in HEK 293T cells transfected with vectors expressing VP16, VP16 mutant, or empty vector (EV), and stimulated with TNF-α. Bottom: Immunoblotting. (b) Top: Amino acid residues 61-98 from HPV16 protein E7 encompassing a ΦXXΦΦ motif containing and preceded by negatively charged residues. Highlighted are two residues mutated to disrupt this motif. Middle: NF-κB-dependent luciferase activity in HEK 293T cells expressing E7 and two mutants as described in (a). Bottom: Immunoblotting. Means + s.d. (n = 4 per condition) are shown. c, Lysates from transfected HEK 293T cells were immunoprecipitated with anti-HA. Immunoblots are representative of two independent experiments. Protein molecular masses in kDa are shown on the left of the blots. Immunoblots of tagged proteins are labelled with the protein name followed the epitope tag antibody in parentheses. When multiple tagged proteins are shown in the same immunoblot, each protein is indicated a red arrowhead. Statistical significance was determined by the Student’s t-test.
