Fig. 2: A high mtDNA mutation burden induces dysfunctional mitochondria and glycolytic addiction without impacting nuclear DNA mutations.

a Schematic illustrating the independently generated TDCLs from NSCLC tumors in KP, PGKP, young, and old animals. b, c The mtDNA mutation burden is represented by SNVs and Indels in PGKP and KP TDCLs. The n for each group is informed in A d,e The nDNA mutation burden is represented by SNVs and Indels. Only the relevant statistics are displayed in the figure. f Growth curve of KP and PGKP TDCLs (Incucyte) derived from young animals. This curve is a combination of four experiments. Supplementary Fig. S11 shows individual growth curves. g The confluence of TDCLs at the end of 80 h. This experiment represents the last time point of the previous growth curves. h The viability of the TDCLs was measured during 13 passages twice. Each point represents one viability test per clone using four KP and eight PGKP clones. i The allograft tumor growth assay was performed using three KP and five PGKP clones in C57/B6 mice. The lines represent the mean values ± SEM. j–l Flow cytometry assessing mitochondrial mass and mitochondrial membrane potential in TDCLs. The graph depicted the data of three independent experiments using four KP and eight PGKP clones m, n OCR and ECAR were measured to evaluate mitochondrial respiration and glycolysis in TDCLs. The graph represents the average of two independent experiments using four KP and eight PGKP clones. Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) and 2-deoxy-glucose (2-DG) were employed in the study. In H, individual sample distributions are represented by dots, while the gray bar indicates the median ± SEM. Boxplot (Min-to-Max) displaying the interquartile range (box), median (line inside the box), and whiskers extending from the minimum to the maximum observed values. Two-tailed t-tests determined P values for G, H, J, K, L, M, and N, while one-way ANOVA with Tukey correction was used for Fig. s B - E. Significance levels are denoted as *≤0.05, **≤0.01, ***≤0.001, and ****≤0.0001. For detailed individual P values, please refer to the Source Data table. The Figures a and i were created in Biorender. Cararo Lopes, E and White, E (2025).