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. 2012 Jul;194(14):3579–3588. doi: 10.1128/JB.00399-12

Fig 1.

Fig 1

Analysis of Legionella entry into macrophage and amoeba hosts. (A) WS treatment reverses defective entry of ΔlpnE Legionella into macrophages. J774 macrophages were infected at 37°C and an MOI of 100 with stationary-phase (Stat) or water stress (WS)-treated bacteria expressing GFP. Means and standard deviations are plotted. On the left, it can be seen that WS treatment of the ΔlpnE strain restores entry to that of parental JR32. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE mutant. For WS bacteria, P = 0.31. On the right, it can be seen that complementation with plasmid pKm14 reverses defective entry of Stat bacteria. For ΔlpnE pKm14 Stat versus ΔlpnE pKm14∷lpnE Stat bacteria, P < 0.001. (B) WS treatment reverses defective entry of ΔlpnE Legionella into amoebae. A. castellanii trophozoites were infected at 28°C and an MOI of 5 with Stat or WS-treated bacteria expressing GFP. Means and standard deviations are plotted. On the left, it can be seen that WS treatment of the ΔlpnE strain restores entry to that of parental JR32. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE mutant. For WS bacteria, P = 0.25 for JR32 versus the ΔlpnE mutant. On the right, it can be seen that complementation with plasmid pKm14 restores parental entry. For JR32 pKm14 Stat versus ΔlpnE pKm14 Stat, P < 0.001. For JR32 pKm14 Stat versus ΔlpnE pKm14∷lpnE Stat, P = 0.73. (C) The lvh locus is not required for WS reversal of defective ΔlpnE mutant entry into macrophages. J774 macrophages were infected at 37°C and an MOI of 100 with Stat or WS-treated bacteria expressing GFP. Means and standard deviations are plotted. WS treatment of the ΔlpnE and ΔlpnE Δlvh strains restores entry to that of parental JR32. For WS-treated bacteria, P = 0.60 for JR32 versus the ΔlpnE mutant and P = 0.65 for JR32 versus the ΔlpnE Δlvh mutant. WS reverses the entry defect in Stat bacteria. P < 0.001 for Stat versus WS-treated ΔlpnE mutant and P < 0.001 for the ΔlpnE Δlvh mutant. (D) The lvh locus is not required for WS reversal of defective ΔlpnE mutant entry into amoebae. A. castellanii trophozoites were infected at 28°C and an MOI of 5 with Stat or WS-treated bacteria expressing GFP. Means and standard deviations are plotted. WS treatment of ΔlpnE and ΔlpnE Δlvh strains restores entry to that of parental JR32. For WS-treated bacteria, P = 0.61 for JR32 versus the ΔlpnE mutant and P = 0.99 for JR32 versus the ΔlpnE Δlvh mutant. WS reverses the entry defect in Stat bacteria. P < 0.001 for Stat versus WS-treated ΔlpnE mutant and P < 0.001 for the ΔlpnE Δlvh mutant. (E) EnhC is required for WS reversal of defective ΔlpnE mutant entry into macrophages. J774 macrophages were infected at 37°C and an MOI of 50 with Stat or WS-treated bacteria expressing GFP. Means and standard deviations are plotted. WS treatment of ΔlpnE and ΔenhC strains reverses the entry defect in Stat bacteria. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE mutant and for JR32 versus the ΔenhC mutant. For Stat versus WS bacteria, P < 0.001 for the ΔlpnE and ΔenhC mutants. WS treatment of the ΔlpnE ΔenhC double mutant fails to reverse the entry defect. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE ΔenhC mutant. For WS bacteria, P < 0.001 for JR32 versus the ΔlpnE ΔenhC mutant. (F) EnhC is required for WS reversal of defective entry of ΔlpnE Legionella into amoebae. A. castellanii trophozoites were infected at 28°C and an MOI of 5 with Stat or WS-treated bacteria expressing GFP. Means and standard deviations are plotted. WS treatment of ΔlpnE and ΔenhC strains restores entry to that of parental JR32. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE mutant and for JR32 versus the ΔenhC mutant. For WS bacteria, P < 0.75 for JR32 versus the ΔlpnE mutant and P = 0.82 for JR32 versus the ΔenhC mutant. WS treatment of the ΔlpnE ΔenhC double mutant fails to reverse the entry defect. For Stat bacteria, P < 0.001 for JR32 versus the ΔlpnE ΔenhC mutant. For WS bacteria, P < 0.001 for JR32 versus the ΔlpnE ΔenhC mutant.