Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Feb 22;8:3469. doi: 10.1038/s41598-018-21795-y

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2018

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/.

PMC Copyright notice

GSK3 is a negative regulator of Fgf21 expression and secretion in brown adipocytes. (a) Kinase inhibitor screen to identify novel regulators of Fgf21 expression in response to β-adrenergic stimulation. Fgf21 expression in mature immortalized brown adipocytes pre-treated with 10 μM kinase inhibitor for 1 h before stimulation with 0.1 μM ISO for an additional 6 h. Unstimulated (blue) and broad spectrum inhibitor-treated cells (red) serve as controls. Green columns show cells pre-treated with GSK3 inhibitors. (b) Fgf21 mRNA levels in mature immortalized brown adipocytes pre-treated with 10 μM of the GSK3 inhibitors SB415286, SB216763 and BIO for 1 h before being stimulated with 0.1 μM ISO for additional 6 h. (c) Secreted FGF21 in cell culture medium from immortalized brown adipocytes pre-treated with 10 μM SB216763 for 1 h before stimulation with 0.1 μM ISO for an additional 24 h. (d) Immunoblot against HA-tag and total GSK3β in immortalized brown adipocytes overexpressing constitutively active (CA) or kinase dead (KD) GSK3β or empty vector. TFIIB serves as loading control. Full-length blots/gels are presented in Supplementary Fig. 2. (e) Expression of differentiation markers (Fabp4, AdipoQ and Cebpα) in immortalized brown adipocytes after ectopic overexpression of GSK3β mutants. (f) Fgf21 expression in immortalized brown adipocytes overexpressing GSK3β mutants or empty vector. Mature adipocytes were treated with 0.1 μM ISO for 6 h. Data represents mean of means +SEM from 3 independent experiments. Statistical significance was determined by two-way ANOVA with repeated measures and Tukey’s post hoc test for multiple comparisons. *p < 0.05 versus H₂O. ^#p < 0.05 versus vehicle/vector control. No statistical tests were applied to panel a.