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. 2017 Aug 16;7:8484. doi: 10.1038/s41598-017-08921-y

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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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Activating effects of WJs for STZ-MSC. (a) Phase contrast images of STZ-MSC. Bar: 100 µm. (b) The average length of minor axis of cells in phase contrast images. All cells in five panels of each STZ-MSC were measured. N = 4, each. **P < 0.01. (c) TEM images of STZ-MSC. White arrows show mitochondrial degeneration. Yellow arrowheads show abnormal dilation of the ER. Bar: 5 µm in upper panels; 500 nm in lower panels. (d) Immunofluorescence staining of STZ-MSC with Ki-67 antibody (red). Nuclei counterstained with DAPI (blue). Bar: 100 µm. (e) Average ratio of Ki-67-positive cells to total cell count in five panels of each STZ-MSC. N = 3, each. *P < 0.05. (f) MTT proliferation assays of STZ-MSC. Absorbance was measured 0, 24 and 48 hours after MTT addition. WJ(−); n = 7, WJ(+); n = 29. *P < 0.05. (g) STZ-MSC scratch assay. The open wound area was measured immediately and 12 hours after performing the cross-scratch. Yellow lines indicate the cell mobilization edge. (h) The ratio of open wound area at 12 hours compared with 0 hours. Five cross-scratch points in each STZ-MSC were measured. N = 3, each. *P < 0.05. (i) Relative expressions of mRNA in STZ-MSC. WJ(−); n = 4, WJ(+); n = 12. *P < 0.05. (j) Western blot analysis of STZ-MSC cultured with 5 lots of WJs (#1–5) or without WJs using anti-BiP, anti-XBP-1, anti-JNK1/3 and β-actin antibodies. The cropped images of immunoblots displayed in the figure and the full-length blots were shown in Supplementary Fig. S14. (k) Western blot analysis of STZ-MSC cultured with 3 lots of WJs (#30, 37, 35) or without WJs using anti-α-SMA and β-actin antibodies. The cropped images of immunoblots displayed in the figure and the full-length blots were shown in Supplementary Fig. S15. (l) Immunofluorescence staining of STZ-MSC with anti-α-SMA antibody (red). Nuclei counterstained with DAPI (blue). Bar: 100 µm. (m) The exosome count in the STZ-MSC supernatant quantified by CD9 ELISA. WJ(−); n = 3, WJ(+); n = 7. *P < 0.05. Values are expressed a mean ± SE.