Figure 6.

Effects on differentiation markers when ALLN treatment is delayed until after acquisition of DNA-binding activity by C/EBPβ. (A) 3T3-L1 preadipocytes were induced to differentiate and after 20 h were treated with ALLN (26 μM). Cell extracts were prepared at the times (Hrs) indicated and subjected to immunoblotting with anti-C/EBPβ, -C/EBPα, -CHOP-10, and -422/aP2 antibodies. (B) Cells were treated or not with ALLN as in A above. At 24 h after treatment, nuclear extracts were prepared and subjected to EMSA which was performed with C/EBP-binding site oligonucleotide (as in Fig. 4B) as probe. (C) Cells were treated or not with ALLN (for 48 h) as in A above. After treatment with ALLN (or not) for 48 h, cells were shifted to regular medium and on day 6 were fixed and cytoplasmic triglyceride stained with oil red O.