ppk2 is required for M. tuberculosis virulence in the murine model. (A) Separate groups of mice were aerosol infected with the wild-type, ppk2::Tn, and ppk2::Tn Comp strains, and the number of CFU in the lungs of mice were determined at various time points postinfection. Values that were significantly different are indicated as follows: *, P < 0.001 comparing the value for the ppk2::Tn mutant with the value for the wild type or ppk2::Tn Comp strain; **, P < 0.001 comparing the value for the ppk2::Tn mutant with the value for the wild type or ppk2::Tn Comp strain. The values for the ppk2::Tn Comp strain and the wild type were significantly different (P = 0.004). Five biological replicate samples were used for each strain in each experiment. (B) Spleen CFU of the wild-type, ppk2::Tn, and ppk2::Tn Comp strains at various time points postinfection. Values that were significantly different (P < 0.05) are indicated by a bar and asterisk. Five biological replicate samples were used for each strain in each experiment. (C) Histological analysis of mouse lungs infected with wild-type, ppk2::Tn, and ppk2::Tn Comp strains at day 28 postinfection. Magnifications, ×40 (a, d, and g), ×200 (b, e, and h), and ×400 (c, f, and i).