Research ArticleMOLECULAR BIOLOGY

Molecular signatures and functional analysis of beige adipocytes induced from in vivo intra-abdominal adipocytes

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Science Advances  11 Jul 2018:
Vol. 4, no. 7, eaar5319
DOI: 10.1126/sciadv.aar5319
  • Fig. 1 Photographs of the anatomical and morphological ultrastructures of different types of fat depots in bat and mouse.

    (A to F) Bat sWAT, aWAT, and iBAT under 30°C (A to C) and 10°C (D to F). (G to L) Mouse aWAT, sWAT, and iBAT under 30°C (G to I) and 10°C (J to L). Scale bars, 0.5 μm (for ultrastructure) and 1 mm (for the dissected tissues in the inset windows).

  • Fig. 2 Analysis of gene expression and markers for bat aWAT and mouse sWAT.

    (A) Genome-wide gene expression profile and hierarchical clustering. (B) MDS of the DEGs from bat and mouse adipocytes. (C) Venn diagram of the DEGs of 10°C versus 30°C groups in bat and mouse iBATs and beige adipocytes. (D) Gene ontology (GO) analysis of the overlapping marker genes in bat and mouse. NADH, reduced form of nicotinamide adenine dinucleotide. (E) Top common markers of beige adipocytes, BAT, and WAT in bat and mouse. *Mouse beige adipocytes and BAT markers. ^Bat beige adipocytes and BAT markers. HA, H. armiger; MM, Mus musculus.

  • Fig. 3 Functional analysis of Uqcrc1 and Letm1 in beige adipocyte differentiation.

    (A) Oil Red O staining of differentiated white adipocyte cultures and beige-like adipocyte cultures. Scale bar, 50 μm. (B) Diameter of lipid droplets was calculated using ImageJ software. (C and D) Expression of Uqcrc1 and Letm1 in 3T3-L1 preadipocytes and experimentally differentiated adipocytes. (E and F) Protein expression of Uqcrc1, Letm1, Ucp1, and Pgc-1α. W and B represent white and beige adipocytes, respectively. (G) Gene expression of Ucp1, Pgc-1α, C/EBPβ, Prdm16, Cd137, Tmem26, Cited1, and Tbx1. n = 3 for all groups. Data are means ± SD for all bar graphs. *P < 0.05; *P < 0.01; ***P < 0.001. Significant analysis was performed by one-sided Student’s t test.

  • Fig. 4 Letm1 function in adipogenesis and sensitivity to insulin concentration.

    (A) Fabp4 expression in 3T3-L1 preadipocytes and differentiated white adipocytes. (B) Oil Red O staining of differentiated white adipocyte cultures and beige-like adipocyte cultures. (C) Fabp4 expression in differentiated white adipocytes from Letm1-depleted preadipocytes treated with increasing insulin concentrations. (D) Oil Red O staining of differentiated white adipocytes treated with increasing insulin concentrations. Scale bars, 5 mm (for the round dishes) and 50 μm (for microscopic Oil Red O staining). (E) Proposed regulatory pathways of Uqcrc1 and Letm1 in the development of beige adipocytes and Letm1 function in adipogenesis. n = 3 for all groups. Data are means ± SD for all bar graphs. *P < 0.05; **P < 0.01. Significant analysis was performed by one-sided Student’s t test.

Supplementary Materials

  • Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/4/7/eaar5319/DC1

    Fig. S1. Clustered profile of all marker genes.

    Fig. S2. Expression levels of six beige marker genes assayed by RT-qPCR.

    Fig. S3. Ponceau staining as loading control in Western blots.

    Table S1. Mapped Ensembl genes and raw read counts from 20 samples.

    Table S2. All beige adipocyte, BAT, and WAT marker genes.

    Table S3. Primers used for qPCR.

    References (38, 39)

  • Supplementary Materials

    The PDF file includes:

    • Fig. S1. Clustered profile of all marker genes.
    • Fig. S2. Expression levels of six beige marker genes assayed by RT-qPCR.
    • Fig. S3. Ponceau staining as loading control in Western blots.
    • Legends for Tables S1 and S2
    • Table S3. Primers used for qPCR.
    • References (38, 39)

    Download PDF

    Other Supplementary Material for this manuscript includes the following:

    • Table S1 (Microsoft Excel format). Mapped Ensembl genes and raw read counts from 20 samples.
    • Table S2 (Microsoft Excel format). All beige adipocyte, BAT, and WAT marker genes.

    Files in this Data Supplement:

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