The x-axis represents different treatment conditions: D M S O, d N s, G e m, and d d C. The y-axis represents mitochondrial D N A copy number or mean fluorescence intensity. There are eight sets of vertical bars, each set corresponding to a specific treatment condition. Each set contains two types of bars: one for wild-type macrophages and one for A k 4 knockout macrophages. The bars for wild-type macrophages are colored in black, while the bars for A k 4 knockout macrophages are colored in red. Graphs A, B, C, F, and H show a fluctuating pattern, while graphs D, E, and G show a decreasing pattern.
Ak4 controls mtDNA synthesis through the deoxynucleotide metabolism in macrophages after Listeria infection. (A–H) WT and Ak4 KO TG-pMacs were pretreated with DMSO, 10 μM dNs (A, C, and F), 5 μM Gem (B, D, and G), or 10 μM ddC (B, E, and H) for 1 h, followed by infection with Listeria at an MOI of 5 for 1 h. Cells were then treated with 250 μg/ml gentamicin, washed with PBS, and maintained in 50 μg/ml gentamicin for 24 h (A and B) or 6 h (C–H) prior to analysis. (A and B) mtDNA copy number in WT and Ak4 KO TG-pMacs was measured by qPCR (n = 4). (C–H) MFI of MitoTracker Green (C–E) and MitoTracker Deep Red (F–H) in Listeria-infected WT and Ak4 KO TG-pMacs was analyzed by flow cytometry (n = 4). mtDNA copy number was normalized to nDNA. 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. nDNA, nuclear DNA; MFI, mean fluorescence intensity.