Figure 4
From: Methylnaltrexone Potentiates the Anti-Angiogenic Effects of mTOR Inhibitors
Analysis of MNTX, temsirolimus and mTOR regulation of VEGF-induced Akt activation in human EC. Panel A: Human EC were either untreated (control) or treated with scramble siRNA, mTOR siRNA, Src siRNA, Rictor siRNA or Akt siRNA for 48 hours, lysates obtained, run on SDS-PAGE and immunoblotted with anti-mTOR (a), anti-Src (b), anti-Rictor (c), anti-Akt (d) or anti-actin (e) antibody. Panel B: Human EC were serum starved for one hour and either untreated (control) or treated with VEGF (100 nM, 5 minutes) with or without pretreatment (1 hour) with 100 nM MNTX, 100 nM temsirolimus, or pretreatment for 48 hours with mTOR siRNA. EC lysates were obtained, run on SDS-PAGE and immunoblotted with anti-pSer473Akt (a), anti-pThr308Akt (b), anti-AKT (c), anti-pThr389 p70 S6K (d) or anti-p70 S6K (e) antibody.