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. 2024 Oct 2;10(40):eadm9801. doi: 10.1126/sciadv.adm9801

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Copyright © 2024 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 1. — (A) Experimental setup for single-molecule fluorescence scanning assays in zero-mode waveguides. (B) Idealized single-molecule fluorescence trace for scanning signal, indicating eIF1 dwell time, τ. (C) Representative trace with scanning signal observed on the wild-type RPL41A mRNA. (D to G) Representative eIF1 dwell-time distributions on wild-type RPL41A mRNA (n = 193 molecules), RPL41A^AAG (n = 143), RPL41A^AAG/Kozak (n = 94), and RPL41A^CUG (n = 254) mRNAs, respectively. The distribution in the 0 to 20 s time domain is expanded to the left of the full-timescale distribution. Distributions from replicate experiments were statistically indistinguishable. Inset schematics indicate distances (in nucleotides) between the +1 nt and the initiation codon.