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. 2016 Aug 3;7(9):624–637. doi: 10.1007/s13238-016-0300-7

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© The Author(s) 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

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Exposure to the HMF counteracts the negative effects of the removal of growth factors on NS growth. Primary cells were seeded in complete medium, EGF-halved (1/2EGF) or bFGF-halved (1/2bFGF) medium, and cultured in the GMF/HMF condition for 6 days. Compared to cells in the complete medium, NSs significantly decreased in size when the concentration of EGF was halved in the GMF but not in the HMF, and the NS size of the three groups in HMF were significantly larger than those in the GMF. Three trials were completed with 18 repeated wells in each trial. Data are shown as mean ± SEM. P values were calculated using a χ² test for NS size distribution comparison. *P < 0.05; ****P < 0.0001; n.s. P ≥ 0.05