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. 2022 Apr 14;13:1995. doi: 10.1038/s41467-022-29603-y

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© The Author(s) 2022

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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Fig. 4 — a Localization of ZCRP-A to the chloroplasts and b localization of ZCRP-B to the cell membrane. YFP fluorescence is false-colored green and chlorophyll autofluorescence is false-colored red. Composite images are stacks of the individual channels c, d differential interference contrast (DIC), e, f yellow fluorescent protein (YFP), and g, h chlorophyll autofluorescence (Chl auto). i Micrographs of Zn-limited wild-type and ZCRP-A knockout (KO) P. tricornutum cells showing morphological differences. For a–h and i, results were validated > 10 times. j Topology predictions from five sub-methods (OCTOPUS, Philius, PolyPhobius, SCAMPI, and SPOCTOPUS), consensus prediction (TOPCONS), and predicted ΔG values for P. tricornutum ZCRP-B generated using the TOPCONS webserver (https://topcons.cbr.su.se/)^27,28. k Extent of Co uptake after 24 h for wild-type (WT), ZCRPA-knockout (KO), and ZCRPA-overexpression (OE) lines of P. tricornutum normalized to fluorescence units (fsu). Data are presented as mean values ± the standard deviation of biological triplicate cultures (n = 3). Individual data points are overlaid as white circles. The extent of Co uptake was found to be significantly larger in the ZCRPA-OE line compared to the wild-type via one-way ANOVA (f(3) = 23.16, p = 0.000268) and post hoc Dunnett test (p = 0.00048).