Fig. 2.

Gas plasma treatment reduced metabolic activity, motility, and viability in urothelial cancer cell lines. (a) Schematic overview of gas plasma treatment performed in 2D in vitro experiments. (b) representative images of resazurin metabolization. (c) Metabolic activity quantification normalized to respective controls indicating IC₅₀ values (box) calculated from non-linear regression of log-transformed x-values (n = 8); graph shows mean. (d) Representative flow cytometry intensity histogram of caspase 3/7 activation. (e) Apoptosis quantification normalized to respective controls (n = 5); heat map shows mean; statistical analysis was performed using one-way analysis of variance (ANOVA) with Dunnett post-hoc testing (*p < 0.05, **p < 0.01; ***p < 0.001). (f) Representative images of DAPI and Mitotracker Red staining. (g-h) Quantification of (g) percent DAPI⁺ cells and (h) mitochondrial membrane potential intensity evaluated until 24 h after gas plasma treatment normalized against the 0 h control (n = 3); graphs show mean. (i) 10–90 % percentile box plots (mean indicated by + ) showing quantification of cellular motility for (I) RT-112, (II) SCaBER, and (III) T24 cells (n = 3 with 5 technical replicates each); statistical analysis was performed using unpaired t-test (***p < 0.001). ns = non-significant. Scale bar = 100 µm (f).