Fig. 2

TRAIL interacts with IER3. a Huh7 cells were co-transfected plasmids encoding Myc-TRAIL and Flag-IER3, after which IER3- or Flag antibody-conjugated beads were used for co-immunoprecipitation (co-IP) analyses of lysates from these transfected cells. Western blotting was then used to detect TRAIL and IER3 in these samples using appropriate antibodies specific for TRAIL, IER3, Flag, or Myc. For reverse co-IP assays, cells were transfected as above, and then lysates were immunoprecipitated using anti-Myc or anti-TRAIL antibodies, after which TRAIL and IER3 levels in precipitates were measured as above. b Interactions between GST-TRAIL and IER3 were demonstrated in a GST pulldown assay. c TRAIL-Myc and IER3-Flag localization within Huh7 cells were assessed by fixing cells 24 h post-transfection, permeabilizing cells, and staining with antibodies specific for TRAIL (green) and IER3 (red), with DAPI (blue) being used to stain nuclei. Cells were then analyzed via confocal microscopy. Scale bars = 10 μm