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. 2017 Oct 31;7:14398. doi: 10.1038/s41598-017-14719-9

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© The Author(s) 2017

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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CAST overexpression significantly reduced macrophage accumulation. (A) Representative immunofluorescent staining of F4/80 in EpiWAT cross-sections from 16 week LFD and HFD fed CAST WT and Tg mice. The nuclei were stained with DAPI (blue) and the F4/80-positive cells (red) are indicated by arrows. Using fluorescent microscopy, F4/80-positive cells were counted from 10 fields at the power of 100x magnification (n = 5). (B) mRNA abundance of F4/80, CD68, CD11c, CD206, MCP-1 and CCR2 genes in EpiWAT from LFD and HFD fed CAST WT and Tg mice were analyzed by qPCR (n = 5). (C) MCP-1 protein accumulation in adipose tissue explant culture media was measured by ELISA. (D-G) WT and CAST-Tg BMDMs were seeded on transwell filters and lower chambers were filled with media containing either vehicle or MCP-1 (100 µg/mL). Cells that migrated through the membrane stained with hematoxylin and were counted from 9 fields at the power of 200x magnification (H) (n = 4). Values are represented as mean ± SEM. *and # denotes P < 0.05 when comparing LFD vs HFD and WT vs Tg respectively (Two-way ANOVA with Holm-Sidak post hoc analysis).