Fig. 3: Analysis of autophagy flux in human primary macrophages infected with MTBC strains of ancient and modern lineages.

Macrophages were infected with the indicated ancient (a) or modern (b) MTBC strains and with H37Rv as a reference strain. After 20 h, cells were either incubated with E64d + PepA to inhibit lysosome activity or left untreated. Autophagy levels were analyzed for LC3 expression by immunoblotting. GAPDH levels were analyzed to normalize the amount of protein loaded (upper panels). LC3-II/GAPDH ratios were quantified by densitometric analysis using the ImageQuant software. Each point value represents the mean ± SD from three independent experiments (lower panels). NI: not infected