Uptake of mycobacteria by AEC and PuM.

<p>Isolated AEC (A and B) and PuM (C and D) were cultured on cover glass at a concentration of 1×10⁵ cells per well as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032125#s4" target="_blank">Material and Methods</a>. After overnight culture in medium without antibiotics, the cells were infected with GFP-BCG at a MOI of 1∶100 (cell∶bacteria) for 4 h at 37°C in RPMI medium without antibiotics. To kill all extracellular remained bacteria, the cells were treated with gentamicin for 1 h, washed 3 times and finally incubated for 72 h in RPMI without antibiotics. After that, the cover glasses with infected cells were fixed, mounted and observed under white light (A and C) and green light (B and D) in a fluorescence microscope. Magnification 1000×.