The KM-plots for survival and lung histology of NSCLC patients. (A and B) The association of TLT-1 expression with reduced survival in NSCLC and patient subgroups. The FP survival curves for TLT-1 expression extracted from various previous microarray studies for all stage NSCLC patients (LUAD), with hazard ratios and P values as specified. (C) The OS KM-plot for early stage (TNM stage 1) on the left and for non-smokers on the right. (D) The beeswarm plots for all NSCLC patients whose survival curves are shown. The red dots are the samples categorized as “high expression,” while the black dots are the “low expression” category for TLT-1, as per the analysis from the used database. (E) H&E-stained cryosections of human NSCLC tumors. Scale bar represents 100 µm. (F and G) The representative images (F) from dual IHC assay on NSCLC cryosections of tumor-rich or tumor-poor samples showing CD8/TLT-1 staining (top panels) or CD4/TLT-1 staining (bottom panels). The CD4 and CD8 stainings are colored brown, while TLT-1 is represented by pink stain. The slides were counterstained by Gill’s hematoxylin (blue). Zoom in on the square insets shown at either corner in yellow boxes (scale bar = 200 μm) for enhanced clarity. The respective stained areas were quantified (G) in tumor areas (excluding stromal regions; four to eight frames per section) by an observer blinded to experimental groups and are presented as mean ± SEM. Significant differences are indicated with asterisks (*, P < 0.05; **, P < 0.01; ***, P < 0.001). The data represent the results of two independent experiments.