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. Author manuscript; available in PMC: 2014 Feb 26.
Published in final edited form as: Sci Transl Med. 2013 Jul 31;5(196):196ra99. doi: 10.1126/scitranslmed.3005747

Fig. 3. Resistance to PI3K inhibition induced by mTORC1 activation.

Fig. 3

(A) Generation of MDA453 and T47D cell lines with acquired resistance to BYL719. (Right) Proliferation of parental and resistant (MDA453R and T47DR) cells in the presence of 1 μM BYL719. (B) Immunoblotting analysis of phosphorylated proteins in parental, MDA453R, and T47DR cell lines after 24 hours of treatment with 1 μM BYL719. (C) Generation of MCF7 cell line with acquired resistance to GDC-0941. (Right) Proliferation of parental and resistant (MCF7R) cells in the presence of 1 μM GDC-0941. (D) Immunoblotting analysis of phosphorylated proteins in MCF7 and MCF7R after 24 hours of treatment with 1 μM GDC-0941. (E) Reduction in the number of reads by individual FRAP1 shRNAs transfected into drug-resistant cells after 7 days of treatment with either BYL719 or GDC-0941 [normalized to dimethyl sulfoxide (DMSO)–treated controls]. (F) Immunoblotting analysis of phosphorylated proteins in MCF7 and T47D cells after knockdown of TSC2 with shTSC2 and treatment with 1 μM BYL719 for 2 hours. (G) Proliferation assay of MCF7 and T47D cells after TSC2 knockdown with shTSC2 in the presence of 1 μM BYL719 for 4 days. Graph shows one representative experiment of two performed. Data are means ± SEM. P value was calculated using two-sided Student's t test.