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. 2012 Jun 20;40(16):8085–8098. doi: 10.1093/nar/gks530

Figure 6.

Figure 6.

Analysis of function of synthetic 18 S rRNAs containing deletions in 5′ ETS or ITS1. (a) Reporter assays of N2a cells cotransfected with a monocistronic reporter construct expressing an optimized firefly luciferase (Fluc2) and various 18 S rRNA constructs as indicated. A schematic representation of the monocistronic construct is shown above. The synthetic rRNAs used in each experiment are indicated below the graph: p18 S.1 contains the full 5′ ETS and ITS1, p18 S.7 contains a deletion of the 3′ region of the 5′ ETS, and p18 S.2 lacks ITS1. Luciferase activities were determined as described in Methods. The results of the various transfections are reported as fold induction of Fluc2, which is luciferase activity over background obtained with WT (pactamycin-sensitive) ribosomes (p18 S.1). (b) Reporter assays of N2a cells transfected with dicistronic reporter constructs expressing Fluc2 and an optimized human Renilla luciferase (hRen). The control construct contained an MCS in the intercistronic region; the other constructs contained either the EMCV or PV IRES. The 18 S rRNA constructs and reporter constructs used in each experiment are indicated below the graph. The results are reported as hRen to Fluc2 ratios normalized to the MCS construct, which has no IRES activity. Error bars represent standard deviations from 3 independent experiments.