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. 2017 Dec 19;7:17785. doi: 10.1038/s41598-017-18138-8

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© The Author(s) 2017

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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Functional analysis of miR-196a and miR-196b in HNSCC-derived cell lines and CAFs. UT-SCC-42B, FaDu cells and CAFs were transfected with either premiR-196a, premiR-196b or non-targeting control, as indicated. (A) Cell proliferation was estimated by Tetrazolium-based MTS assay after 4 days. Data were normalized to the absorbance at day 0 and relative to control-transfected cells. (B) Bar chart showing the relative migration of UT-SCC-42B, FaDu or CAFs transfected with premiR-196a, premiR-196b or non-targeting control. (C) Representative images from the wound healing assays showing the initial scratch (t = 0) area and the residual area at each established final time (5, 15 or 20 h for UT-SCC-42B, FaDu or CAFs, respectively). (D) Bar chart showing the relative invasion of UT-SCC-42B, FaDu or CAFs transfected with premiR-196a, premiR-196b or non-targeting control. (E) Representative images from the 3D invasion assays of CAFs spheroids embedded into a collagen matrix. The invasive area was determined by calculating the difference between the final area (t = 20 h) and the initial area (t = 0) using Image J analysis program. All data were normalized to control-transfected cells and expressed as the mean ± SD of at least three independent experiments performed in quadruplicate. *p < 0.05, **p < 0.01 and ***p < 0.001 by Student t-test.