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. Author manuscript; available in PMC: 2014 Oct 10.
Published in final edited form as: Cell. 2008 Apr 18;133(2):314–327. doi: 10.1016/j.cell.2008.02.030

Figure 6. Phospho-Upf1 Inhibits the Conversion of 40S/[35S]Met-tRNAiMet/mRNA to Translationally Competent 80S/[35S]Met-tRNAiMet/mRNA in RRLs.

Figure 6

BSA, baculovirus-produced FLAG-Upf1(WT), or baculovirus-produced FLAG-Upf1(G495R/G497E) were added to RRLs prior to the addition of charged [35S]Met-tRNAiMet and RLuc mRNA.

(A) [35S]Met was quantitated (colored plot) and superimposed on absorbance profiles of subpolysomes and polysomes that were separated by sucrose gradient centrifugation. The positions of 40S, 60S, and 80S complexes are specified.

(B) Superimposition of the [35S]Met distribution in sucrose gradients of FLAG-Upf1(WT) or FLAG-Upf1(G495R/G497E) as shown in (A).

(C) RT-PCR of RLuc mRNA in sucrose gradients.