Fig 1. The Kp citrate synthase, gltA, interacts indirectly with Lcn2 during lung infection.
(A) A gltA (VK055_1802) mutant was constructed and used to validate InSeq findings. C57BL/6J mice or isogenic Lcn2-/- mice were retropharyngeally inoculated with approximately 1×106 CFU of a 1:1 mix of WT KPPR1 and KPPR1ΔgltA. Lung bacterial burden was measured after 24 hours, and log10 competitive index of the mutant strain compared to the WT strain was calculated for each mouse strain (n = 10, mean displayed, *P < 0.05, ***P < 0.0005, ****P < 0.00005, one-sample t test or Student’s t test). (B) WT KPPR1 and various isogenic mutants were grown in RPMI + 10% (v/v) heat-inactivated resting human serum ± purified recombinant human Lcn2 overnight, then total CFU was enumerated by dilution plating on selective media (n = 3–4, mean displayed ± SEM, ****P < 0.00005, Student’s t test).