Journal:

Article Title: Deletion of the Gene Encoding p60 in Listeria monocytogenes Leads to Abnormal Cell Division and Loss of Actin-Based Motility

doi: 10.1128/IAI.71.6.3473-3484.2003

Figure Lengend Snippet: (A) Immunofluorescence analysis of Caco-2 cells infected with wild-type (wt) L. monocytogenes, the Δiap mutant, or the iap+ revertant for 6 h. Bacteria appear red due to labeling of surface ActA by the anti-ActA antiserum, and cellular F-actin appears green due to FITC-phalloidin. Filamentous actin is randomly distributed around the Δiap bacteria, in contrast to the F-actin tails visible at the poles of the wt and iap+ intracellular bacteria. (B) Transmission electron microscopy of the Δiap mutant in Caco-2 cells 6 h postinfection. Filamentous actin (arrows) is randomly distributed around the bacteria in both panels.

Article Snippet: After fixation with 4% paraformaldehyde for 30 min and quenching with 10 mM glycine for 15 min, bacteria were incubated for 1 h at RT with an anti-ActA antiserum or the monoclonal anti-internalin A antibody (both diluted 1:50 in PBS), washed extensively with PBS, and then incubated with a secondary FITC-labeled antiserum (Dianova) for another hour.

Techniques: Immunofluorescence, Infection, Mutagenesis, Labeling, Transmission Assay, Electron Microscopy