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. 2020 Oct 23;3:612. doi: 10.1038/s42003-020-01348-8

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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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Fig. 3 — a qPCR of endogenous SCD5 mRNA abundance in response to miR222 overexpression and inhibition. n = 3 independent experiments. “***” Indicates statistically significant. b Western blotting of SCD5 protein level in response to miR222 overexpression and inhibition. GAPDH is the internal control. M is the Page Ruler Prestained Protein Ladder 26616 (ThermoFisher Scientific). c Dual luciferase reporter testing of miR222 on SCD5 3′UTR. pRL-TK is the Renilla luciferase expression plasmid used to normalize luciferase activity. n = 3 independent experiments. “***” Indicates statistically significant. d The binding site of miR222 in the SCD5 3′UTR and the introduced mutation. e TG content alteration in response to miR222 overexpression and inhibition in subcutaneous preadipocytes. n = 3 independent experiments. “***” Indicates statistically significant. f Alteration of C16:1/C16:0 and C18:1/C18:0 fatty acid ratio in subcutaneous preadipocytes in response to treatment with miR222 mimics and inhibitor. n = 3 independent experiments. “***” Indicates statistically significant. g TG content in the subcutaneous fat tissue of WT (MSTN^+/+) and MSTN-KO (MSTN^+/−) pigs (n = 3 biologically independent animals). h TG content in the subcutaneous preadipocytes of WT (MSTN^+/+) and MSTN-KO (MSTN^+/−) pigs (n = 3 biologically independent animals). i C16:1/C16:0 and C18:1/C18:0 fatty acid ratio in the subcutaneous fat tissue of WT (MSTN^+/+) and MSTN-KO (MSTN^+/−) pigs (n = 3 biologically independent animals). j C16:1/C16:0 and C18:1/C18:0 fatty acid ratio of subcutaneous preadipocytes of WT (MSTN^+/+) and MSTN-KO (MSTN^+/−) pigs (n = 3 biologically independent animals). k Representative image of Oil Red O staining of the subcutaneous preadipocytes of WT (MSTN^+/+) and MSTN-KO (MSTN^+/−) pigs. Scale bar, 100 μm.