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. 2016 Jul 19;291(37):19274–19286. doi: 10.1074/jbc.M116.740324

FIGURE 6.

FIGURE 6.

The TTF-1 DNA binding domain is required for TTF-1 to inhibit PPFP transcriptional activity. A, PCCL3-EV or PCCL3-PPFP rat thyroid cells expressing either shControl or shTTF-1 to knock down endogenous TTF-1 were transiently co-transfected with a PPFP-responsive human AQP7 promoter- firefly luciferase plasmid, a pRL-TATA Renilla luciferase internal control plasmid, and either pCagen-EV, pCagen-HA-TTF-1 or pCagen-HA-TTF-1HDD. The cells were treated for 24 h with DMSO or pioglitazone followed by assay of luciferases. The y axis represents firefly luciferase normalized to Renilla luciferase. Statistical significance was evaluated with Student's t test, or ANOVA followed Scheffe's test, **, p < 0.01. B, immunoblots against PPARγ (to detect PPFP) and HA tag (to detect HA-TTF-1 and HA-TTF-1HDD) were performed 48 h post cotransfection of the reporter gene assay described in A. Immunoblotting for β-actin was used as a loading control. A–B, data are representative of two independent experiments.