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. 2017 Dec 13;7:17511. doi: 10.1038/s41598-017-17787-z

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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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Substrate stiffness influences β₂ integrin expression and podosome formation in iDCs. (a,b) Cell surface expression of total β₁ integrins (a, n = 6 donors) and total β₂ integrins (b, n = 8 donors) from iDCs conditioned for 6 days on PAA substrates of 2, 12 or 50 kPa, represented as geometric mean fluorescence intensity measured with flow cytometry. Bars represent mean with SEM. Statistical significance was tested with repeated measures ANOVA. (c) Representative widefield images of iDCs conditioned for 6 days on PAA substrates of 2, 12 or 50 kPa and seeded for 3 hours on FN-coated glass. Shown are vinculin (green), actin (red), and the merged image that includes the nucleus (blue). Inserts top left show enlargement of podosomes, indicated in the white square. (d) Fraction of cells forming podosomes from iDCs conditioned for 6 days on PAA substrates of 2, 12 or 50 kPa (n = 4 donors). More than 50 cells per condition were analysed and only cells containing clear actin dots, accompanied by vinculin rings were counted as positive for podosomes. Bars represent mean with SEM. Statistical significance was tested with repeated measures ANOVA. *p < 0.05.