Figure 4.

Mutations in eS25 reduce 40S–IRES complex formation. Filter binding assays were performed 3 min after mixing 1 nM of the indicated purified 40S with 4 nM CrPV IGR IRES to determine the amount of 40S–IRES complex formed. 40S–IRES complexes formed using 40S subunits isolated from wild-type (white bar) or rps25ΔaΔb are re-presented (Figure 1A) here for direct comparison. Purified 40S subunits from rps25ΔaΔb harboring eS25 expressed from a plasmid as the sole source of eS25 in the cell: K33A, R103A, W27A, R58A and R68A are shown for n≥2 biological repeats with P-values indicated *≤0.05, **≤0.01, ***≤0.001 and ****≤0.0001. P-values determined by unpaired t-test.