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. 2020 Jan 14;19:17. doi: 10.1186/s12936-020-3107-1

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. 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 in a credit line to the data.

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Fig. 1 — Three photoschedules were used to generate temporally-staggered cohorts of gametocytes simultaneously perturbed at different ages. Parasites were collected from donor mice at their ZT0, allowing infections in experimental mice to be staggered by 6-hours so that at the same times GMT, all infections could be sampled and treated with TNF or PBS yet, different ages of gametocytes (19-, 25-, and 31-hours-old) could be targeted. By using Per1/2(−/−) mice housed in constant darkness as the experimental hosts, the relevance of gametocyte age was decoupled from the canonical host circadian-clock-controlled rhythms. The ages of focal gametocyte cohorts (labelled “gametocyte age (h)”, defined as hours post RBC invasion) at each sampling event and treatment (in GMT) are highlighted in bold and the ages of the previous (“younger”) and subsequent (“older”) cohorts are illustrated with faint text. Immature gametocytes not yet detectable via microscopy are denoted by “ND”, and gametocytes not yet produced are denoted by “NA”