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. 2019 Sep 12;10(9):676. doi: 10.1038/s41419-019-1931-4

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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. 2 — a–e SMCs cultured in full-serum medium were treated with trehalose (0–100 mM) for indicated time points. Cell proliferation was analyzed by counting the cell numbers (a). Immunoblotting analysis (b, c) shows the effects of trehalose (24 h) on the cell cycle protein cyclin D1 and CDK4. d–e Immunofluorescence images show the proliferative cell marker Ki67 (red) in SMCs treated with vehicle or 100 mM trehalose for 24 h. Nuclei were stained with DAPI. f–j SMCs were transfected with scramble or TFEB cDNA plasmids for 24 h and then cultured in full-serum medium for indicated time points. f Cell number counting. g, h Immunoblotting analysis shows the effects of TFEB overexpression on cyclin D1 and CDK4 in SMCs 48 h after transfection. i, j Immunofluorescence images for Ki67 (red) in SMCs transfected SMC SMCs 48 h after transfection. Scale bar = 50 µm *P < 0.05 (n = 4)