Figure 6

Exogenous treatment with XA induces isoatp4056 expression and A. phagocytophilum burden in tick salivary glands and tick cells. QRT-PCR analysis showing expression of isoatp4056 (A and C) and bacterial loads (B and D) upon treatment with xanthurenic acid (A and B) or xanthurenic acid plus Ro-61-8048 (an inhibitor of XA biosynthesis) at (C and D) different doses in A. phagocytophilum-infected tick cells is shown. Mock controls were treated with the same amount of solvent used for the preparation of XA and the inhibitor. Each circle/square/triangle/inverted triangle represents data from one independent well of the culture plate performed in duplicates. (E) Fluorometer measurements of tick cells infected with GFP-A. phagocytophilum treated with mock (gray circles) or 100 μM XA (black circles) at 510 nm is shown. Each circle represents data from one independent well of the culture plate performed in duplicates. (F) Immunoblotting analysis with anti-GFP antibody showing levels of GFP protein in tick cells infected with GFP-A. phagocytophilum treated with mock or XA (100 μM). Levels of proteins observed on Ponceau stained membrane (used for immunoblotting analysis) serves as a loading control. QRT-PCR analysis showing levels of isoatp4056 (G) or bacterial burden (H) after 24 h post-microinjection in mock- or XA-injected A. phagocytophilum-infected tick salivary glands is shown. Each circle represents data from pair of salivary glands isolated from individual unfed nymphal ticks. P value from non-paired Student’s t test is shown.