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. 2021 May 26;6:187. doi: 10.1038/s41392-021-00579-3

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© The Author(s) 2021

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. 3 — Exosomal S100A4 is a key promoter to enhance metastatic potential. a iTRAQ mass spectrum screening was performed to analyze the protein cargo of exosomes derived from MHCC97-H (HMH-exosomes) and HepG2 exosomes (LMH-exosomes). Proteins were clustered at the 2-fold changes with a p value less than 0.05. The volcano plots revealed a total 116 up-regulated proteins (right quadrant, generally in red, S100A family in black, EIF/EEF family in purple, APO family in blue and PSM family in orange), and yet 43 proteins were found down-regulated (left quadrant, in green). b The results are also expressed as a heat map, from which 116 up-regulated proteins are shown. c Diverse expression abundance of S100A protein family in HCC cell lines. Western blot showed that expressions of S100A4 were the highest in MHCC97-H among HCC cell lines. S100A4 was only highly expressed in exosomes derived from MHCC97-H. d S100A4 knock-down in MHCC97-H results in inhibited migration and invasion ability. Overexpression of S100A4 enhanced migration and invasion in HepG2. e S100A4^rich exosomes significantly promotes migration and invasive abilities of cells. Exosomal S100A4 promotes the migration and invasive abilities of cells in vitro. Exosomes of MHCC97-H cells with S100A4 knockdown were defined as S100A4^low exosomes, ones derived from the scrambled counterparts were defined as S100A4^rich exosomes. S100A4^rich exosomes were used to treat MHCC97-L and HepG2 cells for 24 h before migration and invasion assays while PBS and S100A4^low exosomes were used as negative controls. f Exosomal S100A4 promotes the metastatic potential in vivo. MHCC97-L cells treated with PBS, S100A4^rich exosomes or S100A4^low exosomes were injected in nude mice to form lung metastasis model. All the in vitro assays were conducted three times with three repetitions. Error bars represent the mean ± SD, and the dots represent the value of each experiment; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: no significance. Statistical significance was determined by unpaired t test