Fig. 5

Slug inhibits TR4-TR4RE interaction and TR4 function by suppressing TR4 homodimer formation. a Gel shift assays using ³²P-labeled FATP1-DR1 or PC-DR1. Mock lysate, in vitro translated TR4, and Slug were incubated with ³²P-labeled FATP1-DR1 or PC-DR1, as indicated. b A ChIP assay in 3T3-L1-Slug and 3T3-L1-C cells using an anti-TR4 antibody or normal IgG. The region (− 554 bp to − 341 bp) containing a TR4RE and an E box, and the 3ʹ downstream region (− 143 bp to + 84 bp) lacking both, a TR4RE and an E box of the FATP1 promoter were amplified with immunoprecipitates using PCR. c The effect of TR4 on the interaction between ³⁵S-labeled Slug and purified GST-TR4 protein (GST-TR4-∆C or GST-TR4-FL) was analyzed by a GST pull-down assay. d The effect of Slug on TR4 homodimer formation. In vitro translated ³⁵S-labeled TR4 was incubated with either GST or GST-TR4-FL in the absence or presence of in vitro translated Slug and Slug-2. e and f After 48 h transfection of Slug expression plasmids or SlugmiR plasmids in adipocytes (3T3-L1-TR4 or 3T3-L1-C), mRNA levels of indicated genes (e) and lipid accumulation (f) were determined at day 8 by RT-qPCR and Oil Red O staining, respectively