FIG 7.

The actin nucleator WASH restricts the intracellular replication of L. pneumophila and affects the LCV composition. (A) D. discoideum Ax2 or ΔWASH was infected (MOI, 1) with GFP-producing L. pneumophila JR32(pNT-28), and intracellular replication was assessed by fluorescence analysis. Data show the means and standard deviations for three biological replicates (means of technical replicates each). (B to E) D. discoideum Ax2 or ΔWASH producing calnexin-GFP (pAW16) (B), GFP-Rab1 (pAW7) (C), GFP-Rab7 (pAW9) (D), or GFP-Rab8 (pAW10) (E) was infected (MOI, 5) for 2 h with mPlum-producing virulent L. pneumophila JR32 (pAW14). Shown are representative IFC images (top) and quantifications of IFC colocalization scores (bottom) between GFP and mPlum for >230 cells per sample at the time points postinfection indicated. Data show the means and 95% confidence intervals from one representative experiment out of three independent experiments (***, P < 0.001).