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. 2018 Jan 12;8:583. doi: 10.1038/s41598-017-19073-4

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© The Author(s) 2018

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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IGF-I:IGFBP-3:VN complex stimulates growth of melanoma spheroids in 3D cultures. WM35 and Sk-MEL28 cells were seeded onto 100% Matrigel™ (a,b) or encapsulated in GelMA hydrogel (c,d). On day 10 (Matrigel) or day 21 (GelMA), cells were stained with FDA/PI. TRI 30 = 1 ng/mL VN + 30 ng/mL IGF-I + 90 ng/mL IGFBP-3; TRI 100 = 1 ug/mL VN + 100 ng/mL IGF-I + 300 ng/mL IGFBP-3; TRI 300 = 1 μg/mL VN + 300 ng/mL IGF-I + 900 ng/mL IGFBP-3. Bar graphs represent frequency of spheroid sizes. n = 2 (tested in three wells in each experiment). Asterisks indicate treatments significantly different to VN (*p < 0.05, **p < 0.01). 0% FCS = 3D growth medium without serum. Scale bars indicate 100 μm (matrigel) and 200 μm (GelMA). Error bars indicate SD.