<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

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

Southern blot analysis of <i>met</i> genes using genomic DNA from different <i>Wolbachia</i> infected species.

<p>The blot was hybridized with a probe corresponding to the full length <i>met1</i> (<b>A</b>) and <i>met2</i> (<b>B</b>) genes from <i>w</i>Mel.</p

PCR primers used in the present study.

<p>PCR primers used in the present study.</p

PCR analysis of <i>met2</i> copies in <i>Wolbachia</i> strain <i>w</i>Tei.

<p>PCR was performed using primers met_102F and met_269R (<b>A</b>), met_1024R (<b>B</b>) or TeiB_1024R (<b>C</b>). Primers met_102F and met_269R detect both A-group- and B-group-like <i>met2</i>, met_1024R detects only A-group-like <i>met2</i>, while TeiB_1024R detects only B-group-like <i>met2</i>. M: 100 base pair molecular weight DNA marker (New England Biolabs).</p

Insect lines and <i>Wolbachia</i> strains used in the present study.

<p>Insect lines and <i>Wolbachia</i> strains used in the present study.</p

Phylogenetic tree of <i>Wolbachia</i> based on <i>met</i> gene sequences.

<p>The tree was constructed by Maximum Likelihood analysis. Numbers on the nodes indicate bootstrap values.</p

Distribution of phage methyltransferases in <i>resc</i>+ and <i>resc</i>− <i>Wolbachia</i> strains.

<p>Distribution of phage methyltransferases in <i>resc</i>+ and <i>resc</i>− <i>Wolbachia</i> strains.</p