Fig. 1: High mtDNA mutation burden limits NSCLC growth and malignancy without affecting tumor initiation.

a At the top is a diagram illustrating the generation of the NSCLC conditional model with a high mtDNA mutation burden, PGKP. For breeding strategy, refer to Suppl Fig. S1. Below is a representation of mtDNA sequencing for KP and PGKP animals at various ages (n indicated per animal genotyping). b Accumulation of mtDNA mutations, depicted by single nucleotide variants (SNVs) and Indels, in tissues and conditions presented in panel A. p-values comparing young (p = 0.0001) and old (p = 0.0007) KP and PGKP tumors. Two pup lungs were used as a baseline for both KP and PGKP groups. Three animals were included in the tumor-free groups, and five animals were used for the NSCLC groups. c Survival curve of KP and PGKP, young and old animals. The median survival is illustrated to the right of the survival curve. d In young animals, tumor burden and number, 15 weeks after cancer initiation. H&E shows lung lobes affected by NSCLC. e Wet lung weight after 15 weeks of NSCLC initiation, with accompanying images illustrating normal lungs and NSCLC conditions. Only the relevant statistics are displayed in the figure. f–g Tumor area, number, and wet lung weight after six weeks of cancer initiation. h–j IHC for ki67, cleaved-caspase 3, and TUNEL assay were conducted on PGKP and KP tumors 15 weeks post NSCLC initiation (see Suppl Fig. S3a for representative images). For each group, we used 6 animals, capturing at least 12 images of distinct tumors from each animal. Data is shown as ± SEM. In H-J, individual sample distributions are represented by dots, while the gray bar indicates the median ± SEM. Two-tailed t-tests determined P values for B, D, F, H-J, while one-way ANOVA with Tukey correction was used for Figs. C, E, and G. Log-rank (Mantel-Cox) calculated P values for the survival curve. 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 c were created in Biorender. Cararo Lopes, E and White, E (2025).