Figure 6.

Failure to reduce MPO concentration, ROS production, and necrosis upon MPO inhibition and M. tuberculosis infection in bone-marrow-derived mouse neutrophils in vitro. C3HeB/FeJ bone marrow cells were cultivated for 2 d at the presence of rG-CSF and infected with Mtb at MOI 3. (A) MPO concentrations were quantified by ELISA, (B,C) MPO activity was colorimetrically measured using an artificial substrate (TMB), (D) intracellular ROS production was determined by DHR123 fluorescence using flow cytometry, and (E) necrosis rates were assessed by quantification of extracellular LDH activity. Gating strategy for flow cytometry in Supplementary Figure S5. Depicted is one experiment (A) or two independent experiments pooled in one graph (B–E). Error bars indicate SEM, two-way ANOVA with Sidak’s multiple comparisons test. ***: p ≤ 0.005, ****: p ≤ 0.001.