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. 2016 May 18;6:56. doi: 10.3389/fcimb.2016.00056

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Copyright © 2016 Ozanic, Gobin, Brezovec, Marecic, Trobonjaca, Abu Kwaik and Santic.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Transmission electron micrographs of pulmonary tissues of Balb/c mice. (A) At 2 and 24 h after infection of mice with F. novicida, A. castellanii and/or F. novicida-infected A. castellanii lungs were excised, fixed with glutaraldehyde and processed to electron microscopy. Uninfected mice were used as negative control. Thin black arrows show intact vacuolar membranes, tick black arrow show amoeba and white arrows show bacteria. (B) Quantitative analyses of the intact FCV (Francisella containing vacuole). The integrity of phagosomal membrane was determined by electron microscopy counting at least 100 bacteria for each sample and using following criteria: (a) cytosolic localization of bacteria, (b) vacuolar localization of bacteria- intact vacuoles. ^*p < 0.05.