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. 2019 Dec 20;10:5828. doi: 10.1038/s41467-019-13782-2

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© The Author(s) 2019

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. 4 — a FBP17 expression was silenced in human fibroblast and cells were treated with hypo-osmotic medium (10 min). Trypan blue-labeled cells were scored and the fraction of dead cells was calculated. N = 5 biologically independent samples from 5 independent experiments. Statistical analysis with a two-tailed unpaired t test. *P < 0.05. b FBP17-depleted human fibroblasts were subjected to stretching. Scored as in a. N = 3 biologically independent samples from 3 independent experiments. Statistical analysis with a two-tailed unpaired t test. **P < 0.01. c–g Effect of the indicated treatments in caveolar structures. Quantification of the indicated structures observed by EM of human fibroblasts under hypo-osmotic shock for different time points (in minutes) and after shock release (back to iso-osmotic conditions, 5+5 iso) is shown. N = 45 (iso), 47 (2 hypo), 58 (5 hypo), 43 (10 hypo), and 65 (5+5 iso) biologically independent cells from 3 independent experiments. Statistical analysis with a two-tailed unpaired t test. *P < 0.05; ***P < 0.005. h, i PM tension was measured using magnetic tweezers in different cell lines. Change of the mean tether force after hypo-osmotic shock is shown. N = 12 biologically independent cells from 12 independent experiments (h) and n = 24 (MEF), n = 17 (MEF Cav1 KO), and n = 21 (MEF Cav1 KO FBP17 KO) biologically independent cells from the corresponding independent experiments (i). Statistical analysis with a two-tailed unpaired t test. *P < 0.05; **P < 0.01. j Cells were treated with hypo-osmotic medium and the fraction of trypan blue-labeled cells were scored. N = 7 (MEF), n = 6 (MEF Cav1 KO), and n = 6 (MEF Cav1 KO FBP17 KO) biologically independent experiments. Statistical analysis with a two-tailed unpaired t test. *P < 0.05. k FBP17 tubulation activity is sensitive to stretching. HeLa cells were transfected with GFP-FBP17 and stretched or kept without stretching. The cells were fixed and the amount of FBP17-labeled tubules was scored. N = 224 (unstretched) and n = 226 (stretched) biologically independent cells from 3 independent experiments. Statistical analysis with a two-tailed unpaired t test. *P < 0.05. Data represent mean ± S.E.M. ns non-significant.