Fig. 1. The levels of mtDNA are increased in human and mouse LUAD.
(A) Correlation coefficient and P value for the regression slope between TFAM expression and mtDNA levels in LUAD (14). (B) Linear regression models fitted with gene expression as explanatory and mtDNA levels as response variables. FPKM, fragments per kilobase per million reads. (C) Schematic timeline from tumor induction to collection of the KrasG12D model. (D) Western blot analyses showing the steady-state levels of TFAM in lung homogenates from controls and Kras mice; vinculin (VCL) is used as loading control. (E) Densitometric quantification of TFAM protein levels from Western blots. N = 6; *P = 0.0250. (F) Relative mtDNA levels measured in lung homogenate from controls and Kras mice. N > 8; **P = 0.0058. (G) mtDNA levels measured in laser-captured tumor portions (TUM) and normal lung (NT). Controls: N = 14, Kras: N = 20 ****P < 0.0001. (H) Western blot analyses of TFAM and cytochrome c oxidase subunit 2 (MT-COII) levels in lung mitochondria; voltage-dependent anion channel (VDAC) is used as loading reference. (I) Quantification of TFAM protein levels from Western blots. Controls: N = 12; Kras: N = 9; ns, not significant. (J) Western blot analyses of the steady-state levels of heat shock protein 60 (HSP60), VDAC, and adenosine 5′-triphosphate (ATP) synthase F1 subunit alpha (ATP5A) in lungs; heat shock cognate 70 (HSC70) and Coomassie are used as loading references. (K and L) Quantifications of VDAC and ATP5A levels from Western blots normalized against the loading control. N = 6; *P = 0.0198 and **P = 0.0038. Scatter plots show individual data points and the mean values ± SD. Two-tailed t test was used for statistical analysis of (E), (F), (I), (K), and (L), and one-way analysis of variance (ANOVA) was used for (G). a.u., arbitrary unit.