Fig 6.

Asp14 is presented on the A. phagocytophilum surface. (A) Intact DC bacteria were incubated with trypsin or vehicle control, lysed in RIPA buffer, fractionated by SDS-PAGE, and immunoblotted. Western blots were screened with antiserum targeting Asp14, Asp55, Omp-1A, or APH_0032. Data are representative of at least two experiments with similar results. (B) Live transgenic A. phagocytophilum DC organisms expressing GFP were incubated with preimmune mouse serum, mouse anti-Asp14, or serum recovered from an A. phagocytophilum-infected mouse. Primary antibodies were detected with anti-mouse IgG conjugated to Alexa Fluor 647. Flow cytometry was used to determine the percentage of Alexa Fluor 647- and GFP-positive DC organisms per sample. The fold increase in the percentage of Alexa Fluor 647-positive, GFP-positive DC organisms for each sample relative to preimmune serum is provided. Results presented are the means ± SD for three experiments. Statistically significant (*, P < 0.05) values are indicated.