Figure 3
From: The human papillomavirus E6 protein targets apoptosis-inducing factor (AIF) for degradation
E6 binds to the FAD domain of AIF. (a) List of schematics of AIF and the various constructs used in the study. In all constructs, except full length AIF, AIF was fused with an ubiquitin polypeptide at the N-terminus, and all AIF proteins had an HA-tag at the C-terminus. In some constructs (Δ1, Δ2, Δ3, Δ4, Δ10, Δ11, and Δ12), AIF was fused to GFP at its C-terminus. The results of 16E6 binding to AIF fragments were transferred from (b). A circle (○) indicates binding, and × indicates no binding. MLS: mitochondrial localization sequence; FAD: flavin adenine dinucleotide-binding domain; NADH: nicotinamide adenine dinucleotide-binding domain; C-terminal: C-terminal domain. (b) 293TT cells were co-transfected with p16E6-SBP and AIF or variant expression plasmids by using PEI, and then the proteins in the cell lysates were pulled down with Streptavidin Mag Sepharose and immunoblotted with anti-HA-tag and anti-SBP antibodies. Three independent experiments were performed and one of them is shown. Numbers on the left are molecular masses (in kDa). PD: pulldown; IB: immunoblot.
