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. 2019 May 20;2:190. doi: 10.1038/s42003-019-0443-1

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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. 5 — Early, late, and recycling vesicular compartments in CD and CTR skin fibroblasts. a, b Levels of the EEA1 protein in CD and CTR fibroblasts. Representative WB (a) and densitometry analysis (b) of protein lysates from fibroblasts obtained from patients with CD and controls blotted with anti-EEA1 and anti-tubulin, as a loading control. c Statistical analysis of the fluorescence intensity of EEA1 staining/cell. d Quantitative PCR analysis of the EEA1 mRNA in CD fibroblasts compared to controls. Columns represent the means and bars represent the standard deviations of a representative experiment performed in triplicate. Similar results were obtained from 4 patients and 4 controls. e–g Electron microscopy analysis of fibroblasts from controls and CD patients. Representative images and statistical analysis of fibroblasts from 3 patients and 3 controls. Arrows indicate early endosomes in controls and CD biopsies. Scale bars = 100 nm. h, i Levels of the LAMP2 protein in CD and CTR fibroblasts. Representative WB (h) and densitometry analysis (i) of protein lysates from fibroblasts obtained from patients with CD and controls blotted with anti-LAMP2 antibodies, and anti-tubulin antibodies, as a loading control. j Statistical analysis of fluorescence intensity of LAMP2 staining/cell. k, l Levels of the TfR protein in CD and CTR fibroblasts. Representative WB (k) and densitometry analysis (l) of protein lysates from fibroblasts obtained from patients with CD and controls blotted with anti-TfR antibodies, and anti-tubulin antibodies, as a loading control. m Statistical analysis of fluorescence intensity of TfR staining/cell. Samples from five patients and 5 controls were analyzed in all experiments, unless stated otherwise. Columns represent the means, and bars represent the standard deviations. Student’s t-test, *p < 0.05, ***p < 0.001. n, o TfR levels in the membrane fraction of CD fibroblasts compared with CTR samples with and without P31-43 treatment for 3 h. n WB analysis of proteins from membrane fractions of CTR and CD fibroblasts blotted with anti-TfR and NaK-ATPase antibodies. Representative blots. o Densitometry analysis of WB of samples from 4 patients and 4 controls. Columns represent the means, and bars represent the standard deviations. One-way ANOVA Bonferroni corrected. *p < 0.05