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3.3. TNF-α transiently upregulates Msx2 expression in 3T3-L1 cells
TNF-α transiently upregulates Msx2 expression in 3T3-L1 cells. 3T3-L1 AICAR were incubated with adipogenic differentiation medium in the presence or absence of TNF-α for 8 h. Msx2 mRNA levels were determined using RT-qPCR (A), and protein levels were examined by western blot of nuclear extracts (B). Lamin was used as a loading control. ?p cells under our culture conditions. 3T3-L1 cells were transfected with Msx2 expression plasmids and then cultured for 4 days. Oil Red O staining showed that Msx2 overexpression significantly inhibits lipid accumulation (Supplementary Fig. S2A). Furthermore, Msx2 decreased the expression of C/EBPα, PPARγ, LPL and aP2 mRNAs and the protein levels of C/EBPα and PPARγ (Supplementary Fig. S2B and C). The inhibitory effects of Msx2 on the expression of C/EBPα and PPARγ were significant even at 24 h after adipogenic induction (data not shown). There results suggest that Msx2 is a negative regulator of early adipogenic differentiation in 3T3-L1 cells.
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