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. 2020 Jun 25;10:10348. doi: 10.1038/s41598-020-67056-9

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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 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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Alteration of cell densities or intervention of cytokines and inhibitors on the 3D matrix-induced cell fusion ratio via the cell-matrix and cell-cell adhesions in the Raw264.7 cells. (A) DAPI intensity (DNA content) of 2D and 3D (L, M, and H matrices) cultured Raw264.7 cells with/without IL-4 (20 ng/mL) from FACS assay. The DAPI density corresponding to single nucleus (2n), double nuclei (4n), and triple nuclei (6n) respectively. (B) The bar graphs exhibited the percentage of fused Raw264.7 cells under different doses (0, 0.1, and 1) of cell-matrix adhesion inhibitors (VCN and RGDS) in the different matrices (L: black and H: white) for 7 days, *p < 0.05. (C) The Raw264.7 cells fusion rates were calculated by the fused cell (>3 nuclei) number over total cell number from different Raw264.7 cell densities (2 × 10⁵: white, 2 × 10⁶: black, 2 × 10⁷: gray cells/ml) embedded in the 3D culture (L, M, and H matrices). (D) The Raw 264.7 cell fusion rates with supplemetation of CaCl2 or calcium chelator EGTA.