Figure 1
Validation of downregulation by XL765 of PI3K/mTOR pathway signaling by western blot. (A) Phosphoinositide-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway. 4E-Binding protein 1 (4E-BP1) and S6 kinase (S6K) is a downstream target of mTOR. Solid lines: direct interaction; dotted lines: multistep interaction. (B) Quantification of cell number normalized to control for NHAIDHmut (Dark/light blue) and U87IDHmut (Dark/light purple). (C) Cropped western blots of phosphorylated 4E-BP1, 4E-BP1 and β-actin for NHAIDHmut (left) and U87IDHmut (right) model for control and XL765 treatment. Complete blots can be seen in Supplementary Fig. 1. (D) Quantification of p4E-BP1, 4E-BP1 and their ratio (p4E-BP1/4E-BP1) for NHAIDHmut and U87IDHmut model. (E) Cropped western blots of phosphorylated S6K, S6K and β-actin for NHAIDHmut (left) and U87IDHmut (right) model for control and XL765 treatment. Complete blots can be seen in Supplementary Fig. 2. (F) Quantification of pS6K, S6K and their ratio (pS6K/S6K) for NHAIDHmut and U87IDHmut model. Data are normalized to β-actin level. NHAIDHmut cell were treated with 32 μM XL765 for 72 h and U87IDHmut with 12 μM XL765 for 24 h. DMSO was used as vehicle control (NHAIDHmut: 0.16% for 72 h and U87IDHmut: 0.06% for 24 h). Dark blue/purple bar: Control; Light blue/purple bar: XL765-treated. *p < 0.05, **p < 0.01, ***p < 0.001.
