Extended Data Fig. 9: NPM1 loss affects the translation pathway.

a, b, p-eIF4e(Ser209) staining in SI from Apc^fl/+Kras^G12D/+ and Apc^fl/+Kras^G12D/+Npm1^fl/fl endpoint mice (n = 5/group), and quantification in crypts/tumours. Unpaired two-tailed t tests. c, d, p-eIF4e(Ser209) staining and quantification (crypts) in SI from Npm1^+/+ and Npm1^fl/fl mice four days post-induction (n = 5/group). Unpaired two-tailed t test. e, ³⁵S-methionine incorporation to assess protein synthesis in Apc^fl/flKras^G12D/+ (n = 3) and Apc^fl/flKras^G12D/+Npm1^fl/fl (n = 4) organoids. Data, mean ± SEM; unpaired two-tailed t test. f-h, p-eIF2α(Ser51) staining in SI from Npm1^+/+ and Npm1^fl/fl mice four days post-induction (n = 4/group), and H-score/stain intensity quantification. Two-tailed Mann Whitney test (g); multiple two-sided t-tests with Holm–Sidak correction (h). i, j, Western blot for translation pathway components in epithelial extracts from Apc^fl/fl, Apc^fl/flNpm1^fl/fl, Apc^fl/flKras^G12D/+ and Apc^fl/flKras^G12D/+Npm1^fl/fl mice (n = 4/group), and quantification (j). Data, mean ± SEM. Unpaired two-tailed t tests. k, l, Western blot in Apc^fl/flKras^G12D/+ and Apc^fl/flKras^G12D/+Npm1^fl/fl untreated or DMSO/tunicamycin-treated organoids (n = 3/group), and quantification (l). Data, mean ± SEM. Unpaired two-tailed t tests. m, n, Western blot for puromycin incorporation in Apc^fl/flKras^G12D/+ and Apc^fl/flKras^G12D/+Npm1^fl/fl ± DMSO/ISRIB-treated organoids (n = 3/group), and quantification of boxed areas (n). Cyclohexamide used as negative control. Data, mean ± SEM. One-way ANOVA with Tukey’s test. o, BrdU staining in SI from Apc^fl/flNpm1^fl/fl mice ±vehicle/trazodone treatment (n = 4/group). p, BrdU+ cell quantification in SI half-crypts from groups in (o), compared to untreated Apc^fl/flNpm1^+/+ (n = 5) and Apc^fl/flNpm1^fl/fl (n = 5) mice (also presented in Fig. 2c). Data, mean ± SEM. One-way ANOVA with Tukey’s test. q, Survival curves of untreated Apc^fl/+Kras^G12D/+ (n = 17) and Apc^fl/+Kras^G12D/+Npm1^fl/fl (n = 20) (also shown in Fig. 2n), and trazodone-treated mice (Apc^fl/+Kras^G12D/+ n = 7, Apc^fl/+Kras^G12D/+Npm1^fl/fl n = 10). Median survival (days) indicated in brackets. Log-rank (Mantel-Cox) tests. Censored mice denoted as tick marks. r, tumor distribution within equidistant proximal to distal SI and colon sections from untreated Apc^fl/+Kras^G12D/+ (n = 17) and Apc^fl/+Kras^G12D/+Npm1^fl/fl (n = 20) (also shown in Extended Data Fig. 5o) and trazodone-treated Apc^fl/+Kras^G12D/+ (n = 5) and Apc^fl/+Kras^G12D/+Npm1^fl/fl (n = 8) endpoint mice. Two-way ANOVA with Sidak’s test. Colour combinations denote groups compared. P-values in black indicate no significance. Other significant p-values in red. Boxplots boxes extend from 25^th-75^th percentiles, whiskers from minimum to maximum values, and the line denotes the median. Scale bars (a,c,f), 100 μm; (o), 50 μm.

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