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. 2012 Nov 15;8(11):e1003012. doi: 10.1371/journal.ppat.1003012

Figure 4. Legionella RalF capping domain disrupts secretion and the Golgi apparatus.

Figure 4

A) LpRalF capping domain disrupts secretion. Vectors encoding the indicated YFP-tagged L. pneumophila or R. prowazekii RalF constructs were co-transfected into CHO cells along with a plasmid encoding SEAP. Alkaline phosphatase secretion from CHO cells was plotted as the ratio of SEAP in the culture medium to cell-associated SEAP (Secretion Index). Vector alone (YFP) served as a negative control. These data are from at least 3 independent experiments done in triplicate. The results are normalized so the cells expressing the empty plasmid have a secretion value of 100% (* P<0.001). B) Fluorescence microscopy of Hela cells ectopically expressing YFP alone, YFP-LpRalF1–374, YFP-LpRalF1–201 or YFP-LpRalF192–374. Cells were fixed 24 hours after transfection, then stained with anti-GM130 antibody (red). Arrows indicate disrupted Golgi, arrowheads indicate intact Golgi. Bar = 25 µm. C) Golgi disruption was quantified in cells expressing YFP alone, YFP-LpRalF1–374, YFP-LpRalF1–201 or YFP-LpRalF192–374. These data were obtained from three independent experiments. Standard deviations are represented.