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. 2021 Jul 9;12:4217. doi: 10.1038/s41467-021-24445-6

Fig. 3. Thirty-five percent of 5′-UTR mutations across the spectrum of human prostate cancer functionally impact transcription or translation.

Fig. 3

a Per-gene percentages of distinct barcodes associated with an exact match to an expected 5′-UTR sequence by PacBio long-read sequencing. All distinct 5′-UTR sequences were observed by long-read sequencing and linked to an average of 236 distinct 30-bp barcodes. For each gene, the percentage of barcodes associated with an exactly matching 5′-UTR are plotted as black vertical bars (correctly synthesized). Genes are ordered by increasing 5′-UTR length from left to right, and the average rate of exactly matching barcodes is marked by a horizontal dashed line at 85%. A smoothed fit, using loess regression, of percentage matching vs rank order of length is shown in red. b Correlation of normalized read counts per WT and mutated 5′-UTR in each technical and biological replicate for each PLUMAGE DNA sample. Three biological replicates were analyzed for each cell line (293T and PC3). Pearson correlation coefficient was calculated to determine significance and was found to be r > 0.99 for all samples (all p values = 0.00001). c Correlation of normalized read counts per WT and mutated 5′-UTR in each technical and biological replicate for each PLUMAGE total mRNA sample. Three biological replicates were analyzed for each cell line (293T and PC3). Pearson correlation coefficient was calculated to determine significance and was found to be r > 0.8 for all samples (all p values = 0.00001). d Correlation of normalized read counts per WT and mutated 5′-UTR in each technical and biological replicate for each PLUMAGE polysome-bound mRNA sample. Three biological replicates were analyzed for each cell line (293T and PC3). Pearson correlation coefficient was calculated to determine significance and was found to be r > 0.89 for all samples (all p values < 0.0001). e Proportion of all 5′-UTR mutations assayed by PLUMAGE that showed a significant (FDR < 0.1, in pink) change in mRNA transcript or translation levels. f 5′-UTR mutations (190 mutations) that significantly change gene expression affect important cancer-related pathways by KEGG pathway analysis (Fisher’s hypergeometric test, FDR < 0.05). Source data are provided as a Source data file.