Fig. 2: Transcriptomics and immunofluorescence after constriction indicate increased invasiveness and evidence for stem cell-like states.

A MDS of the two groups control (X1, X2, X3 CTRL) and squeezed (X4, X5, X3 SQZD) analysed in triplicates, where X and Y axes represent dimensions that capture the largest differences between samples based on their gene expression profiles. B Corresponding Venn diagram showing unique and overlapping genes between the two groups. C Volcano plot indicating significantly differentially expressed mRNAs between SQZD and CTRL. The grey horizontal line shows P-value cut-off (*p < 0.01) and the vertical dashed lines indicate up/down-regulated genes (<−1.5 and >1.5fold change). Statistical significance was determined using the edgeR exact test (two-sided), with Benjamini-Hochberg correction for multiple comparisons. D Heatmap of the differentially expressed genes. E Bar graph displaying the number of genes involved in the GO biological processes and KEGG pathways associated with metabolic reprogramming (orange), tumorigenicity and metastasis (pink), cell-matrix interactions (green) and cell cycle (purple). Ticks in the gene number x axis are every 5 genes. F Representative fluorescence images of melanoma cancer cells, seeded on glass before (CTRL) and 4 hours after been squeezed (SQZD) into the microfluidic device, for biomarkers relative to stemness characteristic, e.g., CD44, CD271, ABCB5, PRDM14. G Bar graph showing the fold change of fluorescence signal of stemness-related biomarkers for SQZD cells compared to CTRL cells. The results are expressed as the mean ± SEM from five independent experiments. Source data are provided as a Source Data file.