Extended Data Figure 6: RapaLink-1 has a prolonged intracellular half-life in wild-type mTOR cells.
From: Overcoming mTOR resistance mutations with a new-generation mTOR inhibitor
a, MCF-7 F2039S cells were treated with different concentrations of rapamycin, MLN0128, combination treatment or RapaLink-1 for 4 h, at which time the cells were collected. Immunoblot analyses were performed on mTOR signalling. b, MCF-7 cells were treated for 4 h with either DMSO control, 30 nM of rapamycin, 30 nM of MLN0128, a combination of 30 nM of both or 30 nM of RapaLink-1 for 4 h, at which time the treatments were washed out three times with PBS and fresh media was re-added for the indicated times. Immunoblot analyses were performed on mTOR effectors. c, MCF-7 cells were treated with 10 nM of RapaLink-1 and collected at the indicated times. Immunoblot analyses were performed as described earlier. All experiments were repeated at least three times. d, Mice bearing MCF-7 xenograft tumours were treated with one single dose of vehicle or RapaLink-1 (1.5 mg kg−1), tumours were collected at different days after treatment as indicated. Immunoblot analyses were performed on mTOR effectors. e, The weight of the mice treated in the efficacy study shown in f is reported here. f, Mice bearing MCF-7 xenograft tumours were treated as described in Fig. 4c (n = 5 for each group). The results were reported as percentage tumour volume ± s.d.
