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. 2018 Aug 29;9(9):868. doi: 10.1038/s41419-018-0915-0

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© The Author(s) 2018

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. 3 — a and b Conditioned medium (CM) and total cell lysate were collected to determine the secreted SFRP1 level in CM as well as the cytosolic level of SFRP1 and Rab37. Immunoblot showing SFRP1 secretion was decreased in CM derived from Rab37 knockdown (shRab37#1 or shRab37#2) cells (a), while higher secretion level of SFRP1 in CM derived from Rab37^WT or Rab37^Q89L cells compared to vector control (EV) or Rab37^T43N expressing cells (b). c Rab37-specific vesicles were isolated by IP with Flag-tagged antibody in cells. Immunoblot confirmed that SFRP1 proteins were enriched in Rab37-specific vesicles isolated from Rab37^WT or Rab37^Q89L cells. d Confocal microscopy images of Rab37 (Red), SFRP1 (Green) and nucleus staining (Blue) in cells. Insets show magnification of the boxed area in the merge panel. Scale bar: 10 μm. e The dynamics of Rab37-regulated SFRP1 trafficking in live cells. Selected frames from time-lapse images of Vector, Rab37^WT, Rab37^Q89L, and Rab37^T43N cells expressing GFP-tagged SFRP1. Enlarged images of the boxed areas from time-lapse movies with time intervals in seconds are shown. Triangle indicates vertical movement of GFP-SFRP1 and arrow indicates horizontal movement of GFP-SFRP1. Scale bar: 20 μm. (see also Movies S1, S2, S3 and S4)