Figure 8.

TEAD4 dynamically binds to CtBP2 and YAP during adipogenesis. A–D, endogenous co-IP using TEAD4 and CtBP2 antibodies in stably overexpressing TEAD4 3T3-L1 cells without adipogenic mixture induction (A), after mixture induction for 4 h (B), after mixture induction for 1 day (C), and after mixture induction for 4 days (D). TEAD4, CtBP2, or IgG antibodies were incubated with cell lysates stably expressing TEAD4 and analyzed by co-IP. The arrowheads indicate the binding positions. IB, immunoblot; MW, molecular weight. E, dosage GST pulldown assay to detect the effect of YAP on the interaction between TEAD4 and CtBP2. MBP-CtBP2 and dosage-increased MBP-YAP were pulled down by GST-TEAD4YBD, followed by Coomassie Brilliant Blue staining. F, co-IP assay to detect YAP Ser-94 site function and its antagonism effect with CtBP2 for TEAD4 binding. The indicated plasmids were transfected into 293T cells and analyzed by co-IP. The arrowhead indicates the IgG heavy chain, and the asterisk denotes the nonspecific band. G, model showing that the transcription factor TEAD4 cooperates with its cofactors VGLL4 and CtBP2 to repress adipogenesis during 3T3-L1 differentiation. As 3T3-L1 cells undergo adipogenesis, TEAD4 is highly expressed, and then VGLL4 mediates TEAD4's recruitment of CtBP2 to form a ternary complex and bind to PPARγ/Adipoq/KLF8 promotor regions to inhibit their activities, resulting in inhibition of adipogenesis.