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. 2020 Apr 9;18:58. doi: 10.1186/s12951-020-00610-9

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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. 4 — The expression of the RBDV protein and the in vitro cytotoxicity assay. a pRBDV or pIgG1 Fc were transfected into B16-F10 cells and BALB/3T3 cells. The black or white circles mean transfections of LPPC/pRBDV/RBDV/PEG or LPPC/pIgG1 Fc/RBDV/PEG in B16-F10 cells, respectively. The black or white squares mean transfections of LPPC/pRBDV/RBDV/PEG or LPPC/pIgG1 Fc/RBDV/PEG in BALB/3T3 cells, respectively. The cell culture media were analysed at 0, 6, 12, 24, and 48 h for the amount of protein expression by ELISA. LPPC, which encapsulated with RBDV, were all complexed by PEG. The data represent the mean ± SD (n = 3). Significant differences were evaluated by ANOVA with the Games-Howell test and labelled as *P < 0.05. b NK-92MI cells (effector cells) mixed with B16-F10 cells (target cells) at an E/T ratio of 1:1 were co-incubated with IgG1 Fc, RBDV-IgG1 Fc, or VEGF plus RBDV-IgG1 Fc for 5 h, and the cytotoxic activity was determined by MTT assay. The data represent the mean ± SD (n = 6). Significant differences were evaluated by ANOVA with the Bonferroni test and labelled as *P < 0.05 compared with other groups. c Complement mixed with B16-F10 cells was co-incubated with IgG1 Fc or RBDV-IgG1 Fc, and the cytotoxic activity was determined by MTT assay. The data represent the mean ± SD (n = 4). Significant differences were evaluated by ANOVA with the Bonferroni test and labelled as *P < 0.05 compared with other groups