Figure 2.

Aptamer clusters are genuine binders of eIF4e. (a) Representative members of the top 12 clusters of aptamers selected. (b) Relative abundance of each aptamer family as a percentage of all reads over increasing rounds of selection. (c,d) 6% polyacrylamide EMSA of the selected aptamers against eIF4e and BSA at 2 µmol/l of aptamer and 15 µmol/l of (c) protein or (d) stated concentration of protein. Aptamers were stained with SyBr Gold after electrophoresis. The images are composite images put together as the aptamers were ran across several polyacrylamide gels. Original gel images can be found in Supplementary Figure S8. (e) 150 nmoles of eIF4e was incubated with 150 nmoles of each aptamer or water then incubated with m7-GTP-Sepharose beads containing ~500 nmoles of m7-GTP. Half the beads and half the supernatant was then run on a polyacrylamide gel and Coomassie stained.