Journal: Antimicrobial Agents and Chemotherapy

Article Title: Lipopolysaccharide-Binding Alkylpolyamine DS-96 Inhibits Chlamydia trachomatis Infection by Blocking Attachment and Entry

doi: 10.1128/AAC.02391-14

Figure Lengend Snippet: Pretreatment of EBs with DS-96 results in reduction of anti-LOS staining. Purified EBs were treated with DS-96 (8 μM) or DMSO and stained for LOS (A) or MOMP (B) following methanol fixation. Samples were counterstained with DAPI to visualize the total number of bacteria.

Article Snippet: Bacteria were allowed to settle and adhere to the bottom of the plate at room temperature for 1 h. The liquid was aspirated, and bacterial samples were fixed with 100% methanol for 10 min. For immunofluorescence staining, one set of samples was stained with monoclonal anti-LOS antibody (1683; ViroStat) diluted 1:500 in PBS at room temperature for 1 h, washed with PBS, incubated with secondary goat anti-mouse Alexa Fluor 488 (Invitrogen, Carlsbad, CA), and counterstained with 1 μg/ml DAPI in PBS for 5 min. A parallel set of samples was stained with primary goat anti-MOMP antibody (1621; ViroStat) diluted 1:500 in PBS for an hour, washed with PBS, incubated with secondary donkey anti-goat Alexa Fluor 568 (Invitrogen, Carlsbad, CA) at 1:1,000 dilution for an hour, and stained for 5 min with 1 μg/ml DAPI in PBS.

Techniques: Staining, Purification