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. 2020 Oct 13;10:17095. doi: 10.1038/s41598-020-74105-w

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© The Author(s) 2020

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 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/.

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In vivo conjugation of tHBcAg-SC VLPs with P24-ST. (A) Phenotype of leaves infiltrated with tHBcAg-SC and tHBcAg-SC + P24-ST. Severe necrosis was caused by tHBcAg-SC as shown before. Co-expression of P24-ST substantially mitigated this effect indicating successful conjugation of VLPs and P24. (B) Duplicate Western blots of protein samples from leaves infiltrated with tHBcAg-SC + P24-ST or P24-ST alone (left: anti-P24, right: anti-HBcAg) after double sucrose cushion. Expected size of unconjugated P24 is 24 kDa, expected size after conjugation to tHBcAg is ~ 80 kDa. S = supernatant, 25 = 25% sucrose fraction, 70 = 70% sucrose fraction. C + = tHBcAg-SC + GFP-ST as positive control. Blots cropped for clarity, for uncropped blots see Supplementary Fig. S5.