Extended Data Figure 4: RapaLink-1 requires FKBP12 for binding to the mTOR FRB domain.
From: Overcoming mTOR resistance mutations with a new-generation mTOR inhibitor
a, Dose-dependent cell growth inhibition curves of the MCF-7 parental cell line treated with rapamycin, MLN0128, a combination of rapamycin and MLN0128, or RapaLink-1. The cell growth was determined as described in Fig. 1d. b, mTOR–Flag wild type and variants were transfected into 293H cells. The mTORC1 complex was isolated, and an in vitro competition assay in the presence of FKBP12 was performed as described in Fig. 2b. c, MCF-7 cells were treated with either DMSO, RapaLink-1 (10 nM), FK506 (10 μM), or a combination of both for 24 h, at which time the cells were collected. Immunoblot analyses were performed on mTOR signalling. All experiments were repeated at least three times.
