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. 2020 Nov 3;11:465. doi: 10.1186/s13287-020-01989-w

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — miR-214-3p inhibits TNFα-induced inflammatory SMC differentiation from AdSPCs. a–c Passage 3–8 AdSPCs were cultured in SMC differentiation medium in the absence (vehicle (Veh)) or presence of 50 ng/ml TNFα for 4 days to allow for inflammatory SMC (iSMC) differentiation. Total RNA, protein and conditioned culture medium were harvested and subjected to RT-qPCR (a), Western blot (b) and ELISA analyses (c), respectively. d–f miR-214-3p over-expression rescued mature/contractile SMC marker expressions and inhibited TNFα-induced inflammatory cytokine gene expression. AdSPCs were differentiated into iSMCs (DMEM containing 5 ng/ml TGFβ1 and 50 ng/ml TNFα) for 4 days, then transfected with miR-214-3p mimics (miR-214) or negative control (miR ctrl) and cultured for 48 h in the same culture medium. Total RNA, proteins and conditioned culture medium were harvested and subjected to RT-qPCR (d), Western blot (e) and ELISA (f) analysis, respectively. The data presented here are representative (top panels in b and e) or mean ± S.E.M. (a, c, d, f and bottom panels in b and e) of five or six independent experiments (n = 5 or 6). *P < 0.05 (vs vehicle/control); ^#P < 0.05 (TGFβ1/TNFα vs TGFβ1 or TNFα/miR-214 vs TNFα/miR ctrl) (one-way ANOVA with a post hoc test of Tukey’s analysis). miR-214 indicates miR-214-3p