FIG 3.
Enrichment of uhpC-mutant L. pneumophila following serial infection of BMMs treated with 2DG. (a) Strategy for serial passaging of LP02 ΔflaA lux L. pneumophila in 2DG-treated BMMs. L.p., Legionella pneumophila; p.i., postinfection. (b) Table of mutations in uhpC and corresponding predicted amino acid (AA) changes in UhpC protein observed in independent clones of LP02 ΔflaA lux L. pneumophila with the ability to replicate in 2DG-treated BMMs. SNV, single nucleotide variant. (c) LP02 ΔflaA lux (starting strain before the serial enrichment experiment) and LP02 ΔflaA uhpCΔT1074 lux replication in BMMs with or without 0.5 mM 2DG as measured by light emission (RLU). (d) LP02 ΔflaA lux and LP02 ΔflaA uhpCΔT1074 lux replication in BMMs with or without 0.5 mM 2DG as measured by CFU. ***, P < 0.001; ns, not significant (comparing growth in the presence of 2DG to that with no stimulation by two-way ANOVA). Data are representative of at least two independent experiments. Symbols and error bars represent means ± the standard deviations from at least three technical replicates.