Presence of GlmU_Mtb is obligatory for the survival of <i>Mtb</i> in the host.

<p><b>(A)</b> Confocal microscopy images of THP1 cells infected with <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> in the presence or absence of ATc as indicated were taken 48 h post infection. Bacteria were stained with FITC (green) and lysosomes were stained with Lyso Tracker red DND 99 (red). Scale bars, 10 μm. <b>(B)</b> Quantification of percentage co-localization of bacterium and lysosomes in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005235#ppat.1005235.g003" target="_blank">Fig 3A</a>. n = 10 to 15. <b>(C)</b> PMA differentiated THP1 cells were infected with <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> and either ATc (400 ng/ml) or the vehicle were added at 0 h. CFUs were enumerated at different time points post infection. The experiment was performed in triplicates and error bars represent s.e.m. <b>(D)</b> Guinea pigs were infected with 150–200 bacilli CFUs of <i>Rv</i> or <i>Rv</i>∆<i>glmU</i> and Dox was provided in the water as indicated. Guinea pigs (n = 2) were sacrificed on day 1 and homogenates from the lungs were plated in triplicates to determine the implantation. Guinea pigs (6 guinea pigs /group) were sacrificed four weeks post infection and CFUs were determined in the lung homogenates and results were plotted with log₁₀ /lung on the Y-axis and samples on the X-axis. At four weeks post infection mean CFUs for <i>Rv</i> +Dox or <i>Rv</i>∆<i>glmU</i> +Dox and <i>Rv</i>∆<i>glmU</i>–Dox were 5.2, 0 and 5.11 on log₁₀ scale.***<i>p</i><0.0001, two tailed non parametric <i>t</i>-test, error bars represent s.e.m. <b>(E)</b> Overall pathology (upper panel) and histopathology (lower panel, HE, 40X) of the infected guinea pig lungs four weeks post infection. Both <i>Rv</i> +Dox and <i>Rv</i>∆<i>glmU</i> -Dox were having prominent granuloma and necrotic lesions in the central and epithelioid cells and lymphocytes around it. Guinea pigs infected with <i>Rv</i>∆<i>glmU</i> +Dox were showing normal lung parenchyma with clear alveolar spaces. G = Granuloma, AS = Alveolar Space. <b>(F)</b> Granuloma scores from histopathology results.