M.tb culture and infection

M.tb (strain H37Rv) was grown to log phase at 37 °C in Middlebrook 7H9 broth (BD Biosciences), supplemented with 0.5% (v/v) glycerol, and 0.05% (v/v) Tween 80 (Sigma Aldrich) and 10% albumin-dextrose-catalase. Mice were infected with H37Rv via aerosol route using a Middlebrook airborne infection apparatus (Glas-Col) with an infective dose of ∼100 viable bacilli per lung. Colonies forming units (CFU) in tissue homogenates were quantified using Middlebrook 7H11 Agar (BD Biosciences) supplemented with oleic acid-albumin dextrose catalase and 0.5% glycerol.