Skip to main content
. 2018 Jul 25;200(16):e00176-18. doi: 10.1128/JB.00176-18

FIG 2.

FIG 2

2DG restricts L. pneumophila replication in BMMs independent of Atg5, and perturbation of macrophage glycolysis via other inhibitors or glucose substitution does not affect intracellular replication of L. pneumophila in macrophages. (a) LP02 ΔflaA lux L. pneumophila replication measured by light emission (relative light units [RLU]) detected via luminometer (left) or CFU (right) in unstimulated BMMs (no stim) or in the presence of 0.2 mM 2DG (MOI = 0.01). (b) LP02 ΔflaA lux L. pneumophila replication (RLU and CFU) in Atg5+/+ BMMs (derived from LysMCre+/+ C57BL/6 mice) or Atg5−/− BMMs (derived from LysMCre+/+ Atg5fl/fl C57BL/6 mice) with or without 0.2 mM 2DG (MOI = 0.01). (c) LP02 ΔflaA L. pneumophila replication in broth with or without 0.2 mM 2DG from a starting OD of 1.0 as measured by OD (left) and CFU (right). (d) LP02 ΔflaA (pJB908) (thyA+) lux L. pneumophila replication in A. castellanii with or without 0.2 mM 2DG as measured by light emission (RLU). **, P < 0.01; ***, P < 0.001; ns, not significant (comparing growth in the presence of 2DG to that with no stimulation by two-way ANOVA). Symbols and error bars represent means ± the standard deviations from at least three technical replicates. (e) LP02 ΔflaA lux L. pneumophila replication (RLU) in WT BMMs with or without 1.0 mM sodium fluoride (NaF; MOI = 0.05). (f) LP02 ΔflaA lux L. pneumophila replication (RLU) in WT BMMs with or without 1.25 mM sodium oxamate (NaO; MOI = 0.05). (g) LP02 ΔflaA lux L. pneumophila replication (RLU) in WT BMMs in culture conditions containing 11.11 mM glucose or 11.11 mM galactose in the absence of glucose or in the absence of any sugar (no sugar). In each graph, no curves differ significantly (ns, not significant) as assessed by two-way ANOVA. Data are representative of at least two independent experiments.