Fig. 8

4E-BP1^T37/46 phosphorylation and p62 accumulation correlate and together predict the overall survival of breast cancer patients. a p4E-BP1^T37/46 and p62 tissue microarray (TMA) evaluation using IRS scoring. TMAs of paraffin-embedded breast cancer patient samples (n = 1120) were immunostained for p4E-BP1^T37/46 and p62. Shown are representative immunostaining patterns indicating the intensity of staining (IS) and the percentage of positive cells (PS), graded from 0 (none) to 3 (strong) for IS and from <10% (1) to >80% (4) for PS. The IRS-oriented score for p4E-BP1^T37/46 and p62 was determined by multiplying IS and PS. Scale bars: 200 μM. b p4E-BP1^T37/46 and cytoplasmic p62 levels were evaluated using IRS scoring, calculated with VM Slide Explorer 2.2. Cutoff values (IRS Score 5) for classifying low (IRS < 5) and high (IRS > 5) 4E-BP1^T37/46 and p62 expression were determined via Cutoff Finder. The correlation between p4E-BP1^T37/46 and p62 is illustrated across indicated breast cancer subtypes (HR⁺/Her2⁻ (n = 688), TNBC (n = 178) and Her2⁺ (n = 254)). P-values were determined using a two-sided Fisher’s exact test, with significance defined as <0.05. c Kaplan–Meier survival analysis for indicated levels of p4E-BP1^T37/46 and p62 (low: IRS < 5 or high: IRS > 5). Shown is the OS of HR⁺/Her2⁻ breast cancer patients (n = 687) according to p4E-BP1^T37/46 and p62 levels. Significance was determined using a log-rank test: p = 0.050 (across all 4 arms), p = 0.007 (pairwise comparison of double-low (green) versus double-high (red) arms)