Fig. 2From: Distance-dependent inhibition of translation initiation by downstream out-of-frame AUGs is consistent with a Brownian ratchet process of ribosome scanningContext-dependent inhibitory effects on protein synthesis by proximal out-of-frame dATGs. A, B The average GFP intensities (dots) and the 95% confidence intervals (error bars) of Duo variants, grouped by the sequence contexts of the aATG or dATG (letters in green or red, respectively). The number of Duo variants drawn in each panel (n) is shown in its top-right corner (two biological replicates combined). The numbers of A, G, C, and T context Solo variants are 341, 383, 719, and 362, respectively. C The dual-frame reporter experiment to confirm the competition between aAUG and dAUG as the translation initiation site. The reporter is composed of a modified GFP gene in frame 0 (six stop codons were mutated in the +1 frame of its coding sequence, labeled as GFP*), sequences encoding a 2A self-cleaving peptide in frame +1, and a dTomato gene in frame +1. The fluorescence intensities of each dual-frame construct were normalized by the respective control construct as shown at the bottom. The normalized fluorescence intensities of individual replicates are shown by red or green dots and the mean is shown by the black line. P values were given by t-testsBack to article page