The image contains multiple graphs, including bar graphs and line graphs, depicting the relationship between mitochondrial D N A synthesis and macrophage anti-bacterial activity. The bar graphs labeled A and B show the mitochondrial D N A copy number and mean fluorescence intensity of MitoTracker Green in Listeria-infected macrophages over time. Both graphs show a general increasing pattern. Part C shows the protein levels of P g c-1 alpha, T f a m, and beta-actin in infected macrophages. Additional bar graphs labeled D to I display the effects of different treatments on mitochondrial D N A copy number and bacterial loads in macrophages infected with Listeria or Salmonella. The x-axes represent time or treatment conditions, while the y-axes represent mitochondrial D N A copy number, fluorescence intensity, protein levels, or colony-forming units. The graphs indicate that mitochondrial biogenesis is crucial for macrophage anti-bacterial activity, with significant increases in mitochondrial D N A copy number and mass observed after infection. Treatments with deoxynucleosides, Gemcitabine, or 2 prime,3 prime-dideoxycytidine affect both mitochondrial D N A synthesis and bacterial loads. All graphs except graphs A and B show a fluctuating pattern.
mtDNA synthesis is crucial for macrophage antibacterial activity. TG-pMacs or human THP-1 macrophages were infected with Listeria at a MOI of 5 or Salmonella at an MOI of 10 for 0, 2, 6, 12, and 24 h. (A) The mtDNA copy number in Listeria-infected TG-pMacs was determined by qPCR (n = 4). (B) Mean fluorescence intensity (MFI) of MitoTracker Green in Listeria-infected TG-pMacs was analyzed by flow cytometry (n = 4). (C) Pgc-1α and Tfam protein in Listeria-infected TG-pMacs were analyzed by western blotting. (D) mtDNA copy number in dNs-, Gem-, or ddC-treated TG-pMac with indicated concentration after Listeria infection was determined by qPCR (n = 4). (E and F) Intracellular bacterial loads in dNs-, Gem-, or ddC-treated TG-pMacs after infection with Listeria at an MOI of 5 (E) or Salmonella at an MOI of 10 (F) were assessed by plating cell lysates onto TSA plates and counting CFUs at 24 h after plating (n = 4). (G) THP-1 macrophages were infected with Listeria for the indicated time. mtDNA copy number in Listeria-infected THP-1 macrophages was determined by qPCR (n = 4). (H and I) Intracellular bacterial load in dNs-, Gem-, or ddC-treated THP-1 macrophages after infection with Listeria at an MOI of 5 (H) or Salmonella at an MOI of 10 (I) was assessed by plating cell lysates onto TSA plates and counting CFUs at 24 h after plating (n = 4). mtDNA copy number was normalized to nuclear DNA (nDNA). Protein expression levels were normalized to β-actin. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Data are representative of two independent experiments, and each point represents data from one mouse with two technical repeats. Source data are available for this figure: SourceData F1.