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. 2018 Apr 9;8:108. doi: 10.3389/fcimb.2018.00108

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Copyright © 2018 Watkins, Andrews, Wynn, Barisch, King and Johnston.

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) and the copyright owner 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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Exocytosis of Cryptococcus neoformans from Dictyostelium is dependent on WASH and the actin cytoskeleton. (A) Heat killed S. cerevisiae are not exocytosed from WASH null D. discoideum. Example from 12 h time lapse imaging of heat killed S. cerevisiae in WASH null D. discoideum representative of 60 phagosomes containing heat killed S. cerevisiae from three independent experiments (images start at 500 min after phagocytosis). (B) C. neoformans are exocytosed from WASH null D. discoideum. Example from 12 h time lapse imaging representative of 62 phagosomes containing C. neoformans from three independent experiments (images start at 200 min after phagocytosis). Scale bars 5 μm. (C) Quantification of the percentage of exocytosis of C. neoformans and heat killed S. cerevisiae from WASH null D. discoideum from three independent 12 h time lapses. (D) Frequency histogram of combined 35 transit times measured from three independent 12 h time lapses. (E) Exocytosis of heat killed S. cerevisiae from wild type Ax2 but not WASH null or latrunculin A treated Ax2 D. discoideum. Quantification of the percentage of exocytosis from three independent 12 h time lapses. Total of 60 phagosomes were analyzed from each condition. P-values are Fishers test. (F) Exocytosis of C. neoformans from wild type Ax2, WASH null, and latrunculin A treated D. discoideum. Quantification of the percentage of exocytosis from three independent 12 h time lapses. Total of 60 phagosomes were analyzed from each condition. P-values are Fishers test. (G) Transit times of C. neoformans through WASH null and latrunculin A treated Ax2 D. discoideum are not significantly different. P-values are Mann-Whitney test.