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. 2020 Nov 27;3:717. doi: 10.1038/s42003-020-01449-4

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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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Fig. 1 — a Immunofluorescence images of actin cytoskeleton (green), nuclei (cyan) and vinculin (magenta) as a marker of focal adhesions (FA). MSCs were cultured for 3 h, onto functionalized substrates (FN-coated), with serum-depleted and borax (PLLA-B2%, PLLA-B5%) presence in the culture medium. The same experiment was performed after NaBC1 silencing using esiRNAs (esiRNA^NaBC1). Universal negative control esiRNA (esiRNA^NC) was used as a transfection efficiency control. The PLLA-B2% and PLLA-B5% substrates presented clear focal adhesions as a result of vinculin staining that strongly diminished after NaBC1 silencing (see inset magnifications). Scale bar 25 µm. b Image analysis quantification of different parameters related to FA. Total FA number and FA area before and after NaBC1 silencing. Borax induces more and bigger FAs, effect that is reverted after NaBC1 silencing. Twenty images/condition from three different biological replicas were analyzed. The data represented in graphs correspond to n ≥ 11. Statistics are shown as mean ± standard deviation. Data were analyzed by an ordinary one-way ANOVA test and corrected for multiple comparisons using Dunnett analysis (P = 0.05). ***p < 0.001, ****p < 0.0001.