Figure 2.
Effect of tiRNAs on EMCV IRES-mediated translation initiaton.
A. Synthetic 5′-tiRNAAla does not inhibit EMCV IRES-driven translation. Uncapped (dark bars) or capped (light bars) pF/R bicistronic transcripts were translated in RRL. Left panel: The relative ratio of Firefly to Renilla counts. Means and standard deviations are from 3–4 independent experiments (*-p <0.05, compared to no RNA and three control RNAs (ctrl-1,-2,-3) (Tables S1 and S2, Student’s t-test, n=3–4). Right panel: counts derived from EMCV IRES-driven translation of Renilla ORF relative to the no RNA control (100%). For actual luciferase counts, see Table S2.
B. Synthetic 5′-tiRNAAla inhibits translation of EMCV IRES UA7 variant. Uncapped monocistronic transcripts encoding Renilla luciferase under control of different EMCV IRES variants (WT EMCV-UA6 (upper panel) and EMCV-UA7 (lower panel)) were translated in RRL in the presence of control RNAs (ctrl-1,-2,-3), 5′-tiRNAAla or 3′-tiRNAAla. Means and standard deviations are from three independent experiments *-p <0.05, compared to no RNA and three control RNAs (ctrl-1,-2,-3) (Table SI, Student’s t-test, n=3). See also Figure S2. For actual luciferase counts, see Table S2.
