The image contains multiple graphs and a Western blot analysis. One vertical bar graph (A) shows the mitochondrial D N A (m t D N A) copy number in Salmonella-infected thioglycollate-elicited peritoneal macrophages (T G-p M a c s) over time, with significant increases noted at various hours post-infection. Another vertical bar graph (B) displays the mean fluorescence intensity (M F I) of MitoTracker Green in Salmonella-infected T G-p M a c s, indicating changes in mitochondrial mass. The graph shows a fluctuating pattern. A Western blot (C) illustrates the protein levels of P g c-1 alpha, T f a m, and alpha-Tubulin in Listeria-infected or I L-4 slash I L-13-treated bone marrow-derived macrophages (B M D Ms) at different time points; P g c-1 alpha shows highest number of bands, T f a m shows fewer bands than P g c-1 alpha, and alpha-Tubulin shows fewer bands than T f a m. Another Western blot (D) shows i N o s protein expressions in T G-p M a cs treated with varying concentrations of deoxynucleosides (d Ns) following Listeria infection. In the resulting banding pattern, i N O S shows 130 k D bands, while alpha-Tubulin shows 57 k D bands. Two vertical bar graphs in part (E) present the levels of I L-6 and T N F productions in d Ns-treated T G-pMacs with Listeria infection. The graphs show an increasing trend. (F) depicts a bar graph, which shows the m t D N A copy number in T G-pMacs treated with d Ns, Gem, or d d C at different concentrations. The graph shows a fluctuating pattern. Another vertical bar graph (G) shows the m t D N A copy number in Salmonella-infected T H P-1 macrophages over time. The graph shows a general increasing pattern.
mtDNA copy number increases during Salmonella infection, and the expression of iNOS, IL-6, and TNF-α correlates with mtDNA copy number following Listeria infection. TG-pMacs or BMDMs were infected with Listeria at a MOI of 5 or Salmonella at an MOI of 10 for the indicated time. (A) mtDNA copy number in Salmonella-infected TG-pMacs was determined by qPCR (n = 4). (B) Mean fluorescence intensity (MFI) of MitoTracker Green in Salmonella-infected TG-pMacs was analyzed by flow cytometry (n = 4). (C) Pgc-1α and Tfam protein levels in Listeria-infected or 20 ng/ml of IL-4/IL-13–treated BMDMs were analyzed by western blotting. (D) iNos protein expressions in 0, 1, and 10 μM dNs-treated TG-pMacs with Listeria infection were analyzed by western blotting. (E) IL-6 and TNFα productions in 0, 1, and 10 μM dNs-treated TG-pMacs with Listeria infection were measured by ELISA (n = 4). (F) mtDNA copy number in dNs-, Gem-, or ddC-treated TG-pMac with indicated concentration after Salmonella infection was determined by qPCR (n = 4). (G) mtDNA copy number in Salmonella-infected THP-1 macrophages was determined by qPCR (n = 4). mtDNA copy number was normalized to nuclear DNA (nDNA). Protein expression levels were normalized to α-tubulin. 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 FS1.