Fig. 1: Prolonged mitotic arrest leads to ribosome redistribution toward the 5′ UTR. | Nature Communications

Fig. 1: Prolonged mitotic arrest leads to ribosome redistribution toward the 5′ UTR.

From: Upstream open reading frame translation enhances immunogenic peptide presentation in mitotically arrested cancer cells

Fig. 1

a Representative propidium iodide (PI) staining of U-2 OS cells treated with nocodazole (0.5 µM, 16 h). The bar represents the percentage of cells in G2/M phase. Data from one experiment (n = 1). b Metagene profiles of ribosome-protected fragments (RPFs) in proliferating (blue) and mitotically arrested (green) U-2 OS cells (nocodazole, 0.5 µM, 16 h). Inset magnifies the 5′ UTR. Data from biologically independent experiments (n = 3). c Quantification of RPFs distribution in the 5′ UTR and 3′ UTR of U-2 OS cells treated with vehicle or nocodazole (0.5 µM, 16 h). Mean ± SD from biologically independent experiments (n = 3); P values by two-tailed unpaired t-test. d Heatmap of Pearson correlation coefficients between gene RPKM in U-2 OS cells treated with vehicle (control), BI2536 (0.1 µM), nocodazole (0.5 µM), taxol (1 µM), or STLC (5 µM) for 16 h. e, f Metagene profiles (e) and quantification (f) of RPFs in U-2 OS cells treated as in (d) for 16 h. Inset magnifies the 5′ UTR. Data from one experiment (n = 1). g, h, Metagene profiles (g) and quantification (h) of RPFs in U-2 OS cells treated with vehicle, nocodazole (0.5 µM, 16 h), or nocodazole (0.5 µM, 16 h) combined with torin1 (250 nM, 2 h). Inset magnifies the 5′ UTR. Data from one experiment (n = 1). i Schematic of the run-off elongation experiment in U-2 OS cells. j Metagene profiles of RPFs in U-2 OS cells treated with vehicle or STLC (5 µM, 16 h) and harvested as described in (i). uTIS, upstream translation initiation site. Data from one experiment (n = 1). k Violin plots showing the trimmed mean of M values (TMM) distribution of uTIS in U-2 OS cells treated as in (i). Each point represents a TMM-normalized count at a predicted ORF site. Violin width indicates point density; center line is the median; upper and lower bounds are the 75th and 25th percentiles; whiskers represent minimum and maximum values. P value by one-sided Fisher test (no correction for multiple testing). Data from one experiment (n = 1). Source data are provided as a Source Data file.

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