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. 2012 Jan 24;7(1):e30831. doi: 10.1371/journal.pone.0030831

Figure 1. SBM inhibits adipocyte differentiation of 3T3-L1 cells.

Figure 1

Two-day postconfluent 3T3-L1 preadipocytes (day 0) were treated with the indicated concentrations of SBM and was repleted every 2 days along with the relevant media cocktail upto day 8. Cells treated with 1X PBS were used as controls. The assays were performed on day 8. (A) Intracellular lipids were stained with Oil Red O. (B) Absorbance was spectrophotometrically determined at 500 nm after Oil Red O staining. (C) Triglyceride (TG) content (per mg protein) was measured with a triglyceride estimation kit (Sigma). (D) Glycerol-3-phosphate dehydrogenase (GPDH) activity (U/mg protein) was measured using a GPDH activity assay. (E) mRNA (RT PCR) and protein expression (western blot) of the adipogenic transcription factors C/EBPα, C/EBPβ, C/EBP-δ, and PPARγ. Results were expressed relative to untreated cells after normalization to TATA binding protein (TBP) and β Actin. (F) mRNA abundance of C/EBPα, C/EBPβ, C/EBPδ, and PPARγ in SBM-treated adipocytes relative to control differentiated adipocytes after normalization to 18S rRNA. Five biological replicates of SBM-treated adipocytes were tested. The results were confirmed by three independent experiments, which were each conducted in triplicate. Data are expressed as the mean ± SD. *P<0.01, **P<0.05 vs. controls.