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. 2020 Feb 28;10:3701. doi: 10.1038/s41598-020-60619-w

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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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SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot (WB) analysis of P21 pellet after TCEP reduction in healthy donors. Pellets P21 (lane 1), P21^TCEP (lane 2) and SN21^TCEP (lane 3) originated from 9 (A,C–H) and 20 ml (B) of urine were loaded in each lane and stained with colloidal Coomassie (A,G). Nitrocellulose membranes were hybridized respectively with anti: podocalyxin (PODXL) (C) and collectrin (TMEM27) (C’); Insulin-like growth factor binding protein 7 (IGFBP-7) (D) and podocin (NPHS2) (D’); Tissue inhibitor of metalloproteinases 2 (TIMP-2) (E) and myosin 9 (MYH9) (E’); Tumor susceptibility gene 101 (TSG101) (F) and human serum albumin (ALB) (F’); Nephrin (NPHS1) (B) and CD9 antigen (CD9) (H). No reducing condition (-DTT) for CD9 WB (H) and respective protein patter gel (G). After the first acquisition the same membranes in panel C (PODXL), D (IGFBP-7), E (TIMP-2) and F (TSG101) were incubated again with anti TMEM 27(C’), anti NPHS2 (D’), anti MYH9 (E’) and ALB (F’).