Figure 3
From: Oncoprotein p28GANK binds to RelA and retains NF-κB in the cytoplasm through nuclear export
p28GANK interacts directly with RelA. (A) p28GANK colocalized with RelA in the cell. 293A cells were transfected with RelA plasmid or in combination with 1 μg/well (top panel) or 2 μg/well (middle panel) GFP-p28GANK. They were then treated with or without LMB and were analyzed by confocal microscopy after staining with anti-RelA antibody, secondary antibody and DAPI. Green, red and blue signals represent the localization of p28GANK, RelA and the nucleus. Yellow signals (merge) indicate colocalization of p28GANK and RelA. (B) Interaction of overexpressed Myc-tagged p28GANK with Flag-tagged RelA in vivo. HEK293 cells were co-transfected as indicated, and immunoprecipitated using Flag or Myc-tagged antibodies from cells. Immunoprecipitates (IPs) were immunoblotted for Myc-tagged p28GANK or Flag-tagged RelA. (C) Interaction of endogenous p28GANK with RelA in vivo. HepG2 cells were immunoprecipitated using p28GANK(lane 2) or RelA (lane 5) antibodies, and anti-mouse IgG as a negative control (lanes 1 and 4). IPs were immunoblotted for endogenous RelA (lanes 1 and 2) or for endogenous p28GANK(lanes 4 and 5) and whole-cell extracts for p28GANK or RelA as indicated.
