The Mixed-Lineage Kinase DLK Is a Key Regulator of 3T3-L1 Adipocyte Differentiation

Background: The mixed-lineage kinase (MLK) family member DLK has been proposed to serve as a regulator of differentiation in various cell types; however, its role in adipogenesis has not been investigated. In this study, we used the 3T3-L1 preadipocyte cell line as a model to examine the function of DLK in adipocyte differentiation. Methods and Findings: Immunoblot analyses and kinase assays performed on 3T3-L1 cells showed that the expression and activity of DLK substantially increase as differentiation occurs. Interestingly, DLK appears crucial for differentiation since its depletion by RNA interference impairs lipid accumulation as well as expression of the master regulators of adipogenesis C/EBP alpha and PPAR gamma 2 at both the mRNA and protein levels. In contrast, neither the expression nor the DNA binding activity of C/EBP beta, an activator for C/EBP alpha and PPAR gamma, is affected by DLK loss. Conclusions: Taken together, these results suggest that DLK is important for expression of mature adipocyte markers and that its action most likely takes place via regulation of C/EBP beta transcriptional activity and/or initiation of C/EBP alpha and PPAR gamma 2 gene transcription.