Loss of either isoform of PLD decreases phagocytosis.

<p>A) 40x brightfield images of BMDM cultured with human IgG-coated beads for 5 min and then fixed following washes to remove unbound beads. B) Graph of phagocytic index for WT, <i>Pld1</i>^−/−, and <i>Pld2</i>^−/− BMDM as well as WT BMDM treated with FIPI (750 nM, pre-treated for 1 hr). Number of beads/cell was calculated as total number of beads/field divided by total number of cells/field. Index was defined as % of beads/cell phagocytosed by untreated WT BMDM. Extracellular (bound but not phagocytosed) beads were visualized by Cy5-anti human IgG immunostaining and excluded from the quantitation. Three fields were quantitated for each condition/day and the experiment was performed at least 5 times with BMDM prepared from different mice. C) Confocal images of WT, <i>Pld1</i>^−/−, and <i>Pld2</i>^−/− BMDM that were cultured with GFP-expressing <i>Yersinia pseudotuberculosis</i> strain IP32777c at an MOI of 20 for 20 min, washed, fixed, permeabilized, and stained with rhodamine phalloidin (red). Images are ones typical of at least 3 experiments. Bar, 25 µm D) Graph of IP32777c uptake. Bacteria/cell was calculated as above for beads/cell. p-values: * = p<0.05, ** = p<0.005, *** = p<0.0001.