Fig. 9.

Effect of Knockdown of Mrf-2 on C/EBPβ DNA-Binding Activity at the Early Stages of Adipogenesis

Adipogenesis was initiated by IBMX, Dex, and Ins after treatment of confluent 3T3-L1 cells with scrambled siRNA or site 3 siMrf-2. Nuclear extracts were prepared at the indicated time points. Panel A, Immunoblot analysis of nuclear extracts (10 μg). Panel B, EMSAs. Nuclear extracts were incubated with a radiolabeled oligonucleotide probe corresponding to the C/EBPβ binding site in the C/EBPα promoter, in the presence or absence of a 100-fold excess of unlabeled probe, anti-C/EBPβ or anti-C/EBPδ antibody (0.4 μg), as indicated. C, Scrambled siRNA treatment; KD, siMrf-2 treatment. The results shown are representative of two independent experiments.