Fig. 6.
Immunodepletion of eIF3-p44 by anti-eIF3-p44 antibody results in the loss of mRNA translation.
Rabbit reticulocyte lysates were incubated without (lane 1) or with protein A–Sepharose beads that had been preincubated with PBS buffer (lane 2), preimmune serum (lane 3), or anti-eIF3-p44 antibody (lane 4) as described in “Materials and methods.” (A) After centrifugation, the proteins in supernatants were separated by SDS-PAGE and immunoblotted with anti-eIF3-p44 or anti-β-actin antibody. β-Actin was used as an internal control. (B) Treated lysates were tested for in vitro translation activity using the luciferasegene as a template. 35S-methionine–labeled luciferase was analyzed by SDS-PAGE and autoradiography.
