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. 2019 Mar 18;9:4799. doi: 10.1038/s41598-019-41264-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Chlamydial structures and their intracellular colocalization with secretory granules in infected KY-2 cells. (a) TEM of non-infected (0 hpi, I and IV) and infected (48 and 72 hpi, MOI 40) KY-2 cells (II, III, V and VI). Arrowheads and asterisks indicate chlamydial/granular remnants inside vacuolar structures and on the cell surface (72 hpi). (b) Immunofluorescence of perforin (red, upper panels) or COPI (red, lower panels) and chlamydia (green) in infected (MOI 40, 48 hpi) and non-infected cells. DNA (blue) was stained with DAPI. Cross-reactivity of anti-perforin and anti-α-COP with chlamydia was checked with isolated/purified bacteria (Supplementary Fig. S11). (c) Western blot of chlHSP60 expression in infected KY-2 cells (MOI 40) in the presence of PP2 (0.1 μM) or U73122 (10 μM). Cell lysates were analysed by Western blots probed for chlHSP60 and β-actin. (d) After densitometric analysis, chlHSP60 signals obtained for infected cells (48 hpi) were set to 1 (left: intracellular; right: supernatant). (c) Depicts cropped blots obtained by each protein evaluation. Full-length blots are shown in the Supplementary Fig. S12.