Extended Data Fig. 3: A. phagocytophilum infection in response to bioenergetic alterations.
From: Bacterial reprogramming of tick metabolism impacts vector fitness and susceptibility to infection
(a) Schematic of microbial infection upon glycolysis or OxPhos inhibitor treatment in vitro. 1 × 106 ISE6 cells were treated with 50 mM 2-deoxy-D-glucose (2-DG), 0.1 µM rotenone (Rot), 0.5 µM antimycin (Anti), 0.5 µM oligomycin (Oligo) or 20 µM 2,4-dinitrophenol (2,4-DNP) for 1 hour prior to infection with A. phagocytophilum at MOI 50. Cells were collected 2 days post-infection for bacterial quantification. Infected cells without inhibitor treatment (-) served as a control. (b) A. phagocytophilum infection quantified by amplification of 16 S rRNA gene (n = 5 for each condition; n represents individual culture wells). (c) Experimental design of inhibitor treatment on A. phagocytophilum infection in vivo. Created with BioRender.com. (d-e) Ticks were injected with either PBS (-, grey) or 2-DG (purple) at indicated amounts prior to feeding on A. phagocytophilum-infected C57BL/6 mice. (d) Tick weight (n = 17, 29, 14 and 21) and (e) A. phagocytophilum infection (n = 7, 17, 7 and 12) were recorded. (f-g) Ticks were injected with either PBS (-, grey) or 0.8 pmol oligomycin (orange) prior to feeding on A. phagocytophilum-infected C57BL/6 mice. (f) Tick weight (n = 27 and 31) and (g) A. phagocytophilum infection (n = 10 and 12) were measured. (b, d-g) Data are representative of two independent experiments with mean ± SEM. Statistical significance was evaluated by (b, d-e) Kruskal–Wallis test followed by Dunn’s multiple comparisons test or (f-g) two-sided Mann-Whitney test. NS=not significant. Significant p values (<0.05) are displayed in the figure.
