Figure 8.

CpG induces macrophage metabolic and functional rewiring in vivo. (A) Mice were treated for two consecutive days with CpG (20 µg i.v.) or vehicle (Lactated Ringers) and spleens were harvested 1-day (1d), 3-days (3d), 1-week (1w), or 2-weeks (2w) later. Spleens were processed for magnetic isolation of F4/80+ macrophages which were used immediately for respective assays. (B) Mitochondrial content was assessed by MitoTracker Green staining. (C) Intracellular ATP content was measured. (D) Relative phagocytic activity was assessed by emitted fluorescence upon phagocytosis of pHrodo-tagged S. aureus bioparticles and measured by flow cytometry. (E) ROS production was determined by relative dihydrorhodamine 123 (DHR 123) fluorescence. Data are shown as mean ± SEM, n = 5-16/group *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by ANOVA followed by Dunnett’s post-hoc multiple comparison test.