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. 2022 May 7;20:219. doi: 10.1186/s12951-022-01443-4

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© The Author(s) 2022

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. 4 — The SPc-delivered ds-MIRNA internalizes into protoplasts and maize through endocytosis. a Differentially expressed genes related to endocytosis in protoplasts. The treated protoplasts were collected after 2 h incubation. The housekeeping gene ZmUBC was used as the reference gene. Triplicate biological replicates were used for each treatment. Different letters on columns indicate significant differences among the treatments of control, ds-MIRNA and ds-MIRNA/SPc complex (Duncan’s multiple range test, P < 0.05). b Differentially expressed genes related to endocytosis in maize. The ds-MIR164b/SPc complex was applied to the root surface of maize every 24 h, and the expression of above endocytosis-related genes was determined on 4 days after the treatment. c Schematic illustration of the ds-MIRNA/SPc complex delivery into cells. The negatively-charged ds-MIRNA can be assembled with positively-charged SPc into ds-MIRNA/SPc complex, which is delivered into cells through endocytosis. The endocytosis-related genes were up-regulated, and the location of encoded proteins is shown