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. Author manuscript; available in PMC: 2015 May 28.
Published in final edited form as: Cancer Discov. 2014 Jun 9;4(9):1062–1073. doi: 10.1158/2159-8290.CD-14-0159

Figure 1. p110α and mTORC1 are Critical for the Proliferation of NF1-Deficient Tumor Cells.

Figure 1

A, S462, 90-8TL, and LN-229 cells were transfected with pooled siRNAs targeting PIK3CA, PIK3CB, PIK3CD, or non-targeting siRNA. Bar graphs, relative change in cell number from day 0 to 96 hours as compared with control cells transfected with the non-targeting siRNA. Data points, triplicate averages ± SD. Immunoblots depict p110α, p110β, and p110δ protein levels 72 hours after transfection with the indicated siRNA. Actin serves as a loading control. *, P < 0.002. B, Immunoblots showing pAKT and pS6 levels in S462 cells following treatment with indicated inhibitors (4 hours; 500 nmol/L). AKT, S6, and actin serve as controls. C, Bar graphs of S462 and 90-8TL cells treated inhibitors as specified. Numbers represent the relative change in cell number from day 0 to 96 hours as compared with vehicle-treated control cells. Data points, triplicate averages ± SD. *, P < 0.04. D, S462 and 90-8TL cells were transfected with pooled siRNAs targeting MTOR, RAPTOR, RICTOR, or non-targeting siRNA. Bar graphs, relative change in cell number from day 0 to 96 hours as compared with control cells transfected with the non-targeting siRNA. Data points, triplicate averages ± SD. Immunoblots show mTOR, Raptor, Rictor, pAKT, and pS6 levels 72 hours after transfection with the indicated siRNA. AKT, S6, and actin levels serve as controls. *, P < 0.02. E, S462 cells were treated with the rapamcyin (Rap) at 100 nmol/L, Torin1 at 250 nmol/L, or MK-2206 (concentration indicated). Bar graph, relative change in cell number from day 0 to 96 hours as compared with vehicle-treated control cells. Data points, triplicate averages ± SD. Immunoblots show pAKT, pAKT, pTSC2, pS6, and 4E-BP1 levels in the presence of the specified inhibitors. AKT, TSC2, S6, and actin serve as controls. *, P < 0.0001. F, S462 cells treated with either rapamycin at 100 nmol/L, MK-2206 at 5 μmol/L, or both drugs together. Bar graphs, relative change in cell number from day 0 to 96 hours as compared with vehicle-treated control cells. Data points, triplicate averages ± SD. p, phosphorylated.