Analysis of ROS formation in <i>Wolbachia</i> -infected and uninfected Aa23 cells.

<p>(A) Flow cytometric analysis of <i>Wolbachia</i> –infected and uninfected Aa23 cells using the fluorescent ROS marker carboxy-H₂DCFDA. Histograms representative of three replicates are shown. The negative control (shaded) consists of unlabeled cells. Test samples (black lines) include: uninfected Aa23 cells (top panel), uninfected Aa23 cells induced to produce ROS using TBHP (middle panel), and infected Aa23 cells (bottom panel). Carboxy-H₂DCFDA positive cells are represented on each histogram. (B) Microscopic analysis of <i>Wolbachia</i>-infected (I) and uninfected (II) Aa23 cells. Hoechst stain was used to label DNA (left panel). Carboxy-H₂DCFDA was used to label ROS (right panel). Bar, 10 µm.</p

<i>Wolbachia</i> stably infects Aa23 cells and can be cured by antibiotic treatment.

<p>(A) PCR analysis using <i>Wolbachia wsp</i> primers (top) and arthropod 28S primers (bottom) of Aa23 cells treated with 10 ug/ml rifampicin for seven passages. Lane L: molecular ladder. Lane 1: stably infected Aa23 cells. Lanes 2 through 8: cells treated with rifampicin for 1, 2, 3, 4, 5, 6, and 7 passages. Lane 9: negative control. (B) Aa23 cells stably infected with <i>Wolbachia</i> (I) and Aa23 cells cured of <i>Wolbachia</i> using rifampicin (II). Bar, 100 µm.</p

2-D Page of <i>Wolbachia</i>-infected and uninfected Aa23 cells.

<p>(A) Identification of proteins unique to <i>Wolbachia-</i>infected Aa23 cells. Approximately 750 ug of protein extract from an Aa23 cell line stably infected with <i>Wolbachia</i> (I) and a parallel cell line cured of a <i>Wolbachia</i> infection (II) were analyzed. Proteins expressed only in the presence of a <i>Wolbachia</i> infection (ID #1–6) are identified. (B) Gel sections showing proteins selected for LC/MS/MS analysis.</p