Fig. 6

Aspartate pathway enzymes are essential during acute and chronic mouse infections. Conditional knockdowns were constructed of two enzymes of the aspartate pathway: homoserine dehydrogenase (ΔthrA-DUC) (a–d) and homoserine transacetylase (ΔmetX-DUC) (e–f). a, e Effect of gene knockdown/protein degradation on growth was tested in vitro in unsupplemented 7H9 OADC with different concentrations of anhydrotetracycline (atc). b, f Mice were aerosol infected with either ΔthrA-DUC3 or Δmet-DUC5 to deliver ~100 CFU/lungs. Groups of mice were switched to doxycycline containing chow (2000 ppm) at day 0 (red square), day 7 (green diamond), day 21 (purple triangle) and day 42 (blue inverted triangle) to induce knockdown. One group (black) was left untreated throughout the whole experiment. At given time points, 4 mice per group were sacrificed and bacterial burden of lungs (b, f) and spleens (d, h) determined by CFU counts of homogenized, serially diluted tissue. Gross lung pathology and Hematoxylin and Eosin (H&E) staining of lungs at day 189 are shown under (c, g). X-axis crosses Y-axis at limit of detection. Error bars show standard deviations from n = 4 mice. **p-value < 0.01, *p-value < 0.05 in Student's t-test. Source data are provided as a Source Data file