Fig. 5

SMAD4 is heterogeneously expressed in gastric cancer and a decreased expression correlates with patient survival (validation cohort). References for immunostaining analysis according to H-score. Staining intensities ranged from 0 (A; nuclear and cytoplasmic negative) to 3+ (D, nuclear and cytoplasmic strong expression) with 1+ (B; nuclear and cytoplasmic weak expression) and 2+ (C; nuclear and cytoplasmic moderate expression) in between. Black-and-white expression of SMAD4 describes tumors with clearly demarcated areas of complete loss of nuclear and cytoplasmic SMAD4 expression next to areas with retained expression (E). Anti-SMAD4 immunostaining, hematoxylin counterstain; × 400 (A–D) and × 100 (E) magnifications. F Kaplan-Meier curves depicting patients’ survival according to SMAD4 status (Q1–3 vs. Q4; for further details see Suppl. Results). Kaplan-Meier curves demonstrating correlations between cytoplasmic SMAD4 (top row) and nuclear (bottom row) loss in tumor cells and overall as well as tumor-specific survival