Fig. 4

HECTD3 interacts with IKKα. a Exogenous HECTD3 interacts with IKKα in HEK293T cells. Flag-HECTD3 and IKKα (left) or Flag-IKKα and HECTD3 (right) plasmids were cotransfected into HEK293T cells. After Flag-tagged HECTD3 and IKKα proteins were immunoprecipitated with Flag-M2 beads, IKKα and HECTD3 were detected by immunoblotting. b FLAG-immunoprecipitation of FLAG-tagged HECTD3 from HUVECs. c Anti-IKKα antibody was used to immunoprecipitate IKKα from HUVECs. d Confocal microscopy images of Flag-HECTD3 and IKKα in HUVECs are shown. Scale bars, 10 μm. e Schematic diagram shows human HECTD3 and its truncation mutants (top). Flag-IKKα and GST-fused HECTD3 truncation mutants were coexpressed in HEK293T cells. By GST pull-down assay (bottom), GST-H DOC specifically pulled down Flag-IKKα. f Schematic diagram shows human IKKα and its truncation mutants (top). Flag-HECTD3 and GST-fused IKKα truncation mutants were coexpressed in HEK293T cells. By the GST pull-down assay (bottom), GST-IKKα SDD specifically pulled down Flag-HECTD3. Data represent three independent experiments for all of the above experiments