Figure 4. Gemin5 fails to detectably interact with eIF4E in co-IP experiments.

293T cells were co-transfected with gemin5 and eIF4E expression constructs as in figure 2 and cytoplasmic lysate was used for IP using the indicated murine antibodies (IgG, α-myc, and α-FLAG). (A) Input and supernatant samples assayed for levels of over-expressed proteins. (B) Each IP was analyzed by western blot with α-eIF4E (left) and α-FLAG (middle) rabbit antibodies, and by silver stain (right). Blots were intentionally over-exposed to assess possible co-IP. Asterisks indicate mouse light antibody chain used in IP reactions that cross reacts with rabbit secondary antibody.