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. 2016 Jul 29;5:22. doi: 10.1186/s40164-016-0044-3

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© The Author(s) 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 6 — Effects of systemic sirolimus treatment on thymocyte caspase activity (Day 4). One representative from three triplicate experiments is shown. Mice received intraperitoneal DMSO (a) or sirolimus (b) on Days 0–3. On Day 4, thymic fragments were collected and incubated with the caspase-3 substrate Ac-DEVD-AMC with and without zVAD (pancaspase inhibitor) as described in “Methods” section. The tissue homogenates were separated on HPLC and analyzed by fluorescence for the released AMC moieties (reflecting intracellular caspase activity, R _t = 3.44 min). The values of AMC peak areas (arbitrary unit ÷ 10³ per mg) are shown. AMC peak areas decreased by ≥78 % in the presence of zVAD, confirming the cleavage was mediated mainly by caspases