Fig. 1: Digital spatial profiling of primary NSCLC and metastasized tumor tissues.

a Schematic of study design and workflow. NSCLC patients with metastases to the brain (n = 44) were represented in four tissue microarray (TMA) blocks (LB-D1 to D4) for digital spatial profiling (DSP) of the whole transcriptome (18,694 genes). Regions-of-interest (ROI) for DSP were annotated based on histology by a pathologist and immunofluorescence staining with the morphological markers PanCK (for epithelial cells), CD45 (for hematopoietic cells), and GFAP (for brain cells). A total of 119 ROIs (average 0.2 mm² each) were analyzed. The figure was created with BioRender. The scale bar is 100 μm. b RNA-sequencing saturation graph showing that none of the ROIs sequenced had counts below 50%. c Normalization of the RNA sequencing data using the third (Q3) quartile count. The limits of the violin plots represent the upper and lower quartiles, whereas the dots indicate the median. (L) = 30 samples, (LB) = 27 samples, TBME = 19 samples, TIME-L = 15 samples, TIME-B = 8 samples, mLN = 13 samples, BC = 7 samples. d Principal component analysis (PCA) of the DSP data. e Uniform Manifold Approximation and Projection (UMAP) analysis. Source data are provided as a Source Data file.