Trafficking of L. pneumophila within <i>Dictyostelium</i> Lacking Functional PI3Ks

<div><p>(A) Confocal laser scanning micrographs of calnexin-GFP–labeled <i>Dictyostelium</i> (green) wild-type Ax3 (WT denotes untreated or treated with 20 μM LY) or Δ<i>PI3K1/2</i> (denoted by ΔPI3K) infected with DsRed-Express–labeled wild-type L. pneumophila (red) for 1 h or 2.5 h, respectively. Representative examples of spacious vacuoles (WT) and tight vacuoles (WT/LY, ΔPI3K) are shown. DNA was stained with DAPI (blue). Bar denotes 2 μm.</p><p>(B) Quantification over time of spacious LCVs in calnexin-GFP–labeled <i>Dictyostelium</i> wild-type (filled squares denote untreated; filled triangles denote LY) or Δ<i>PI3K1/2</i> (open squares) infected with DsRed-Express–labeled wild-type <i>L. pneumophila.</i></p><p>Data represent means and standard deviations of the percentage of spacious vacuoles from 50–200 total vacuoles per time point scored in four independent experiments.</p><p>(C) Confocal laser scanning micrographs of <i>Dictyostelium</i> wild-type or Δ<i>PI3K1/2</i>, infected for 75 min with wild-type L. pneumophila expressing M45-tagged SidC. Infected amoebae were stained with rhodamine-conjugated anti-L. pneumophila antibody (red), FITC-conjugated anti-M45-tag antibody (green), and DAPI (blue), respectively. Bar denotes 2 μm.</p>