Skip to main content
. 2009 Mar 31;4(3):e5070. doi: 10.1371/journal.pone.0005070

Figure 2. Recombinant GST-Sg4E-BP (4E-BP) interacts with endogenous SgIF4E isoforms in sea urchin extracts.

Figure 2

After incubation of the GST alone (lanes 1–2) or the GST-Sg4E-BP protein (lanes 3–4) in extract prepared from unfertilized eggs (UF, lanes 1 and 3) or from 60 minutes post-fertilization embryos (F, lanes 2 and 4), proteins were affinity purified using Gluthatione Sepharose 4B beads, resolved by 15% SDS-PAGE, analysed by immunoblotting and detected by chemiluminescence using an anti-GST antibody (top and intermediate panels) or anti-eIF4E antibody (bottom panels) as described in Materials and Methods. SgIF4E that co-purified with GST-Sg4E-BP (lanes 3 and 4) was compared with the endogenous SgIF4E detected in 10 µg of total protein extracts (corresponding to 0,5% of the volume used for the purification) loaded separately (lanes 5–6).