Figure 7.
Visualizing Single Ribosomes Decoding an mRNA Molecule
(A–D) Analysis of single ribosomes on the Emi1 5′UTR_long reporter mRNA. (A) Representative images of multiple single ribosome translation events of individual mRNAs (upper). ScFv-GFP intensity was quantified over time for the two mRNAs marked by asterisks with the same color (lower). (B) Increase in scFv-GFP fluorescence from single ribosome translation events aligned at the first detectable scFv-GFP signal (n = 35 individual mRNAs in pink and average in black). (C) Steady-state and then abrupt decrease in scFv-GFP fluorescence from single translating ribosomes (n = 35 individual mRNAs [pink] and average [black]). (D) Single ribosome elongation rates (n = 44) (Supplemental Experimental Procedures). Mean ± SD is shown in (D). Scale bar, 2 μm.