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Fig. 2 | BMC Microbiology

Fig. 2

From: Legionella effector AnkX interacts with host nuclear protein PLEKHN1

Fig. 2

Confirmation of AnkX interaction candidates using in vitro bead-based pull-down assay. a Protein-protein interaction was determined using a bead-based co-precipitation assay. AnkX-HaloTag and the GST-tagged bait proteins were produced individually by IVTT. The GST-bait proteins were captured using anti-GST beads, and retention of query proteins was determined by SDS-PAGE. Top panel (pull-down, PD) shows that AnkX-HaloTag, but not HaloTag, was captured by GST-bait-coated beads. Beads coated with GST-CYP4F8 or GST-DBN1 did not capture AnkX-HaloTag. Bottom panel (western blot, WB) shows detection of the GST-bait protein by immunoblot using anti-GST antibody. b Protein-protein interaction was further illustrated using well-NAPPA (wNAPPA). AnkX-HaloTag and GST-bait were produced by IVTT and protein complexes were captured by well-immobilized GST antibody. Wells coated with HaloTag served as a negative control. Retention of AnkX-HaloTag was detected spectroscopically

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