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. 2022 Mar 22;204(4):e00060-22. doi: 10.1128/jb.00060-22

FIG 1.

FIG 1

Characterization of the M. tuberculosis espK::Tn mutant. (A) Schematic of transposon insertion into the espK gene in the esx-1 locus of M. tuberculosis Erdman. (B) Growth comparison of wild-type M. tuberculosis Erdman (WT) and isogenic espK::Tn mutant strain in 7H9 (left) and Sauton’s (right) medium (average of 2 independent experiments; error bars represent the standard deviations). (C) Immunoblots of CF (10 μg/well) and CL total protein (5 μg/well) of wild-type M. tuberculosis Erdman (WT), espK::Tn, and 5′ Tn::pe35 strains cultured 3 days in Sauton’s medium without detergent to examine indicated proteins (representative of 3 independent experiments). (D) Cytotoxicity assay in THP-1 cells infected with wild-type M. tuberculosis Erdman (WT), espA::Tn, and espK::Tn strains at an MOI of 5 (data represent means and standard errors of means from 4 independent experiments with replicate wells; the y axis indicates cytotoxicity values relative to uninfected control; significance in difference calculated using Student's t test). (E) Bacterial burden after 3 and 6 weeks postinfection in lungs (top) and spleens (bottom) of mouse aerosol challenged with wild-type M. tuberculosis Erdman (WT) and espK::Tn strain (each data point indicates the bacterial burden per mouse, with median and standard deviations from 4 mice per group per time point shown; significance in difference calculated using Student's t test; NS, not significant).