Association of MSI2 expression with outcome. (A) Kaplan-Meier survival analysis based on manually measured cytoplasmic or nuclear H-scores, showing significant association with outcome for nuclear H-scores. Patients with higher nuclear H-scores have poorer outcome (P = .0055) but lack association for cytoplasmic H-scores (P = .16); the patients were grouped by H-score into 2 groups, one with H-scores less than the 75th centile (quartiles 1-3) and one with H-scores equal to or above the 75th centile (quartile 4), with the first quartile used for the lower end of gene expression for each gene. (B) Kaplan-Meier survival analyses based on Aperio measured percentage of cells showing total cellular MSI2 positivity. Analyses are shown for 1+, 2+, or 3+ intensity of staining and using either the median or 75th centile to split the patients into 2 groups. Higher positivity for MSI2 was significantly associated with a poorer outcome at 3+ intensity of staining above either the median (P = .0069) or the 75th centile (P < .0001) and for 2+ intensity above the 75th centile; the most significant association was for patients with 3+ staining above or below the 75th centile (P < .0001), with patients in the fourth quartile having poorer outcome. (C) Kaplan-Meier survival analyses based on Aperio measured percentage of cells showing nuclear MSI2 positivity. Analyses are shown for 1+, 2+, or 3+ intensity of staining and using either the median or 75th centile to split the patients into 2 groups. Higher positivity for MSI2 was significantly associated with a poorer outcome at 3+ or 2+ intensity of staining above either the median (P = .0425 and P = .0353 for 3+ and 2+, respectively) or the 75th centile (P < .0001 and P = .0241 for 3+ and 2+, respectively) and for 1+ intensity above the 75th centile (P = .0318). The most significant association was for patients with 3+ staining above or below the 75th centile (P < .0001), with patients in the fourth quartile having poorer outcome.