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. Author manuscript; available in PMC: 2015 Feb 7.
Published in final edited form as: Oncogene. 2013 Sep 23;33(32):4156–4163. doi: 10.1038/onc.2013.397

Figure. 8. Binding of eIF3i to COX-2 cRNA.

Figure. 8

A and B. Pull-down assay. Purified recombinant eIF3i was incubated with biotin-labeled or control unlabeled COX-2 cRNA (A) or pre-incubated with unlabeled probe competitor prior to incubation with labeled probe (B) followed by pull down with Streptavidin-beads and Western blot analysis of eIF3i. C. Electrophoretic Mobility Shift Assay (EMSA). Purified recombinant eIF3i was pre-incubated without or with unlabeled cRNA probe competitors followed by incubation with [32P]-labeled COX-2 cRNA and 14-3-3-σ cRNA. The unbound probes were digested with RNase T1 before separation of eIF3i-bound probe on non-denaturing PAGE and detection by autoradiograph. D. Schematic model of eIF3i function and regulation in colon oncogenesis. It is noteworthy that β-catenin is a known activator of COX-2 expression and, thus, there may be a feed-forward loop that involves eIF3i.