Data on regulation of the gene for the adipocyte-enriched micropeptide Adig/Smaf1 by qPCR analysis and luciferase reporter assay

This article describes qPCR analysis for the Adig/Smaf1 gene in multiple in vitro adipocyte differentiation models including white and brown adipogenesis, cell lines and primary cultures. The article also contains qPCR data for transcript levels of Adig/Smaf1 in a wide panel of murine tissues. Expression of Adig/Smaf1 transcript in white and brown adipose tissue in fasted and refed mice is reported and also data for Adig/Smaf1 transcript expression in genetically obese ob/ob mice. Data on the effects of siRNA-mediated knockdown of Srebp1c on Adig/Smaf1 transcript levels in 3T3-L1 adipocytes are shown. Luciferase reporter assays provide data for regulation of an ~ 2 kb fragment of the 5′ flanking region of Adig/Smaf1 gene by PPARγ/RXRα. This data is related to a research article describing Adig/Smaf1 protein expression, “Expression, regulation and functional assessment of the 80 amino acid Small Adipocyte Factor 1 (Smaf1) protein in adipocytes” (G. Ren, P. Eskandari, S. Wang, C.M. Smas, 2016) [1].


Type of data
Adipogenesis, isolation of adipose tissues and liver for qPCR analysis. Transfection of siRNA and expression vectors in cultured cells for qPCR analysis or luciferase assay.

Experimental features
Adipogenesis of 3T3-L1 preadipocytes and of other cell culture models of in vitro adipocyte differentiation was induced using standard methods. Tissues were harvested from C57BL/6 mice and from genetically obese ob/ob mice. RNA was extracted from cells and tissues by Trizol method, and cDNA was synthesized and utilized for qPCR studies. Hela cells were transfected with an Adig/Smaf1 luciferase reporter construct in the absence or presence of co-transfection of PPARγ and RXRα expression constructs to determine transactivation of the Adig/Smaf1 promoter region using luciferase reporter assay.

Toledo, OH
Data accessibility All data is available within the article.

Value of the data
The Adig/Smaf1 gene encodes an adipose-enriched micropeptide of 80 amino acids [1] and there is a growing interest in the functions of proteins encoded by small open reading frames (sORFs) [2].
The Adig/Smaf1 gene was recently implicated in the regulation of human leptin levels based on GWAS data [3].
The function of Adig/Smaf1 remains unknown, therefore quantitative data on regulation of the Adig/Smaf1 gene may provide insight to its metabolic or other roles.
The data can be compared and contrasted with data for other adipocyte genes or genes induced in fatty liver of obese mice.
The data provides a foundation for additional studies of how PPARγ regulates the Adig/Smaf1 gene.

Data
Here we significantly extend the limited information available to date for transcript expression for Adig/Smaf1 in adipose cells and tissues, previously determined by this laboratory [4] and by others [5]. This includes graphical qPCR data for expression of this gene during preadipocyte to adipocyte conversion using multiple in vitro adipogenesis models and with obesity and nutritional status in murine tissues. We also report data on regulation of Adig/Smaf1 gene expression by transcription factors PPARγ and Srebp1c using siRNA and/or luciferase reporter assay.

Cell culture, in vitro adipocyte differentiation and treatments
Methods for culturing and differentiating the various cell lines utilized in this study have been previously described [1,6,7].

Animal studies
Studies were in strict accord with the guidelines in the Guide for the Care and Use of Laboratory Animals of the NIH. For Figs. 1 and 2 data of Adig/Smaf1 transcript expression in wild type C57BL/6 and ob/ob mice tissues, and for fasting and refeeding, are as described [7,8].

RNA preparation and qPCR
For Figs 1-4, RNA was extracted from cultured cells and mouse tissues, further processed for qPCR, and qPCR carried out and analyzed as previously described [4,7].
Cell lysates were harvested 24 h later and further analyzed as described [8]. Statistical analyses were conducted using single factor ANOVA.

Funding source
Funded in part by NIH, United States grant 5R21DK66055 to C.M. Smas and by a grant from University of Toledo Center for Diabetes and Endocrine Research.