Fig. 1

BIRC5 and RGN are differentially methylated and expressed in NSCLC and tumor-free lung tissues. a Normalized methylation analyses of CpG loci on Human Methylation 450k BeadChip (upper top bar: green—controls, blue—adenocarcinomas, red—squamous cell carcinomas; lower top bar: green—controls, red—tumor samples; heatmap: yellow high, blue low DNA methylation values; mean DNA methylation = 0; the target IDs of the CpG loci, their localization, and functional association are indicated). b Relative gene expression level of BIRC5 and RGN as quantile-normalized expression values of tumor-free lung tissues and matched tumors depicting the mean in the 99.9% confidence interval with error bars (green: controls, blue: adenocarcinomas, red: squamous cell carcinomas). One-way ANOVA and Tukey’s multiple comparison test were used with p ≤ 0.001 (***) regarded significant. c Survival analysis using RNA expression data retrieved from kmplot.com selecting Affymetrix-ID 202094 for BIRC5 and 210751_s for RGN. d Exemplary results from IHC analyses displaying expression of BIRC5 (upper and middle panel) and RGN (lower panel) on the protein level in tumors (left) and tumor-free lung tissues (right) from the same donors (scale bars = 100 μm). Middle panel shows the expression of BIRC5 in tumor-adjacent immune cells. e Correlation of DNA methylation (cg06472116, located on chr. X) vs. gene expression (RGN) in female [left graph] (Pearson’s correlation coef − 0.4127; p value < 1.1550 × 10^–15) and male [right graph] donors (Pearson’s correlation coef − 0.5682; p value < 1.3136 × 10^–44)