Fig. 3.

TET3 regulates TRα1 activity and TRα1 protein level. (A) TET3 regulation of TRα1 activity. HEK293T cells were transfected with TRα1, a luciferase reporter plasmid, and 2 doses of TET3 constructs: TET3N that lost interaction with TR; TET3 and enzymatic dead mutant (TET3mut). Relative luciferase activities were measured 24 h after T3 (10⁻⁸ M) treatment; the triangles represent increasing amount of TET3 constructs. The asterisks indicate the significance of the differences between each condition and TRα1 alone in the presence of T3. (B) TR target gene expression is regulated by TET3 levels in C17.2Sα cells. Expressions of TRα1 target genes in indicated cells treated or not with T3 (10⁻⁸ M) were examined by relative qRT-PCR. Relative induction triggered by T3 in each cell line was presented. The asterisks indicate the significance of the differences between two indicated conditions. (C) Effect of TET3 inhibition on TRα1 protein level in C17.2 cells. TRα1 was detected using an anti-TRα1 antibody in C17.2GSα, C17.2SαC, and C17.2SαKO cells. Streptavidin beads were used to precipitate SBP-TRα1 in C17.2SαC and C17.2SαKO. (D) TRα1 binding to target genes after TET3 KO in C17.2Sα. ChAP of TRα1 on TRE or control (Ctrl) regions of indicated genes. Results are presented as percentage of input.