Aberrant accumulation of PTTG1 induced by a mutated thyroid hormone β receptor inhibits mitotic progression
J. Clin. Invest. Hao Ying, et al. 116:2972 doi:10.1172/JCI28598 [Go to this article.]

Figure 2
Analysis of the interaction of PTTG1 with TRβ1 or PV by GST pull-down assays. (A) [³⁵S]-labeled PTTG1 (20 μl) prepared by in vitro transcription/translation was incubated with agarose beads (lane 2), agarose-GST (lane 3), or GST-TRβ1 (lane 4) as described in Methods. Lane 1 shows the input (1 μl) of [³⁵S]-labeled PTTG1. (B). Binding of truncated [³⁵S]-labeled TRβ1 to GST-PTTG1. Lanes 1–5 show the input of [³⁵S]-labeled TRβ1 and truncated TRβ1 proteins as marked (10% of that in lanes 6–10). Lanes 6 and 7 show the binding of the full-length and truncated (domains C+D+E) TRβ1, respectively. The volumes of programmed lysates were adjusted such that equal amounts of full-length TRβ1, ED41, MD32, KD25, and KP24 protein were used in the binding assay. (C) Schematic representation of full-length and truncated TRβ1 proteins, with domains and boundaries indicated. (D) Binding of [³⁵S]-labeled TRβ1 (10 μl) or [³⁵S]-labeled PV (10 μl) to GST-PTTG1 and GST-PTTG1ΔC in the presence (lanes 6, 8, 10, and 12) and absence (lanes 5, 7, 9, and 11) of T3 (1 μM). Inputs (lanes 1 and 2) were 10% of that used in the binding experiments (lanes 5–12). The binding site of PTTG1 with TRβ1 and PV was located in the aminoterminal domain of PTTG1 (lanes 5–12). (E). Schematic representation of full-length and truncated PTTG1 proteins, with boundaries indicated.