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. 2021 Apr 14;11:8177. doi: 10.1038/s41598-021-87759-x

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© The Author(s) 2021

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 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/.

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SIRT1 promoted NMNAT2 expression during 3T3-L1 differentiation and constitutively inhibited NMNAT1/3 expression. (a) Illustration of the subcellular localization of NMNAT enzymes. (b–d) qPCR data comparing the expression of NMNAT1 (b), NMNAT2 (c) and NMNAT3 (d) at specific time-points during adipogenesis in ShScrambled or SIRT1-depleted (ShSIRT1) (pre)adipocytes (n = 4). (e) Effect of supplementation with 1 mM NMN on total cellular NAD⁺ levels in 3T3-L1 preadipocytes (n = 4). (f) Experimental design to test the effect of NMN supplementation on gene-expression during 3T3-L1 adipogenesis. (g) Representative images of Oil Red O (ORO)-stained ShScrambled or SIRT1-depleted (ShSIRT1) adipocytes with (+NMN) or without (−NMN) supplementation with 1 mM NMN. Scale bar, 100 μm. Images were collected at Day 6 post-differentiation and are representative of 3 independent experiments. For statistical analysis of data shown in (b–d), pair-wise comparisons were made between ShScrambled and ShSIRT1 groups for each individual time-point using the Student’s t test (n = 4). A Student’s t test was used to perform statistical analysis on the dataset shown in (e) (n = 4) (*P < 0.05; **P < 0.01).