*3.2. Reduced NME1 Expression Is Significantly Associated with Poor RFS Irrespective of Histology or Pathologic Stage*

Univariate analysis was performed to discover prognostic factors that affect RFS in early stage NSCLC. RFS was negatively associated with reduced NME1 expression but not with β-catenin overexpression and cisplatin-based adjuvant chemotherapy (Supplementary Table S2). Patients were stratified according to histology and pathologic stage to analyze whether the relationship between RFS and NME1 expression was modified by histology or pathologic stage. RFS was compared between patients with and without reduced NME1 expression in histologic subtypes. Reduced NME1 expression was significantly associated with RFS (*p* < 0.0001; Supplementary Figure S1A): Five-year RFS rate after surgery was 38% for those with reduced NME1 expression and 68% for those without reduced NME1 expression. Reduced NME1 expression had a negative effect on RFS in adenocarcinoma (*p* < 0.0001; Supplementary Figure S1B) and in squamous cell carcinoma (*p* < 0.0001; Supplementary Figure S1C).

The effect of reduced NME1 expression on RFS was further analyzed based on pathologic stage (Figure 2). The number of patients with stage IIIA NSCLC was five, which was too small to analyze RFS. Therefore, patients with stage IIIA NSCLC were combined with those who had stage IIB NSCLC to analyze RFS. Reduced NME1 expression was significantly associated with poor RFS in stage IA (*p* = 0.0005; Figure 2A), stage IB (*p* = 0.001; Figure 2B), and stage IIA (*p* = 0.01; Figure 2C). It was marginally associated with poor RFS in stage IIB–IIIA (*p* = 0.08; Figure 2D). The relationship between β-catenin overexpression and RFS was also analyzed based on pathologic stage and histology. However, no association was found between them.

**Figure 2.** Impact of NME1 on recurrence-free survival (RFS) according to pathologic stages. The effect of reduced NME1 expression on RFS was estimated using the Kaplan–Meier survival curve in 77 patients with stage IA (**A**), 161 with stage IB (**B**), 98 with stage IIA (**C**), and 89 stage IIB–IIIA (**D**). Statistical difference between two survival curves was calculated using the log-rank test.
