Novel Role of NOX in Supporting Aerobic Glycolysis in Cancer Cells with Mitochondrial Dysfunction and as a Potential Target for Cancer Therapy
Figure 1
POLGdn expression led to the depletion of mtDNA-encoded respiratory chain components.
(A) POLGdn-pcDNA4/TO construct and nucleotide sequencing analysis confirming the D1135A mutation. (B) Induction of POLGdn expression by doxycycline. T-Rex293 cells carrying POLGdn construction were incubated with doxycycline at an indicated time point. POLGdn expression was detected by anti-FLAG antibody, while both the endogenous POLG and POLGdn proteins were detected by anti-POLG antibody using Western blot assay. (C) Dramatic decrease of mtDNA by expression of POLGdn. Southern blot assay was used to measure mtDNA content. 10 µg total cellular DNA (including genomic DNA and mtDNA) from each sample was digested with SphI to linealize the circular mtDNA, followed by gel electrophoresis. 32P-labeled mitochondrial COII DNA fragment was used as a probe to detect mtDNA. (D) Assay of mtDNA-encoded COII RNA expression by northern blot analysis. (E) Detection of mitochondrial DNA-encoded COII protein by Western blot assay.