FIG 5.

Recruitment of the SCF ubiquitin ligase components Skp1 and Cul1 beneath attachment sites of L. pneumophila to hMDMs and their subsequent compartmentalization to the LCV. Untreated (A) or cytochalasin D-pretreated (B) hMDMs preloaded with anti-Skp1 or anti-Cul1 antibodies were infected by wild-type (WT) L. pneumophila or the isogenic dotA or ankB mutant for 15 min, followed immediately by fixation and processing for microscopy. Representative confocal microscopy images of infected hMDMs cells show colocalization of Skp1 and Cul1 proteins (red) with the LCV (green) (A) and with attached extracellular L. pneumophila (B). The arrowheads indicate intense colocalization of Skp1 or Cul1 with the WT bacteria. The numbers in the merged images of all panels are quantifications of the frequency of recruitment of Skp1 or Cul1 beneath attached extracellular bacteria. The data represent analyses of 100 infected cells and are representative of three independent experiments.