Skip to main content
. 2017 Jul 24;10:229. doi: 10.3389/fnmol.2017.00229

Figure 2.

Figure 2

Role of Aβ in the activation of Akt and mammalian target of rapamycin complex 1 (mTORC1). (A) SK-N-MC were incubated with Aβ (5 μM) and NAC (1 mM) for 24 h. Phosphorylated Akt (Thr 308 or Ser 473), Akt and β-actin were detected by western blot. n = 6. (B) Cells were incubated with Aβ for 0–48 h. Phosphorylated mTOR (Ser 2448 and Ser 2481), mTOR and β-actin were detected by western blot. n = 3. (C) Cells were treated with Aβ for 24 h. Protein samples were immunoprecipitated by using mTOR antibody-conjugated protein A/G agarose beads. Immunoprecipitation assay was described in “Materials and Methods” section. Raptor, Rictor and mTOR were detected by western blot. n = 3. (D) Cells were pretreated with LY294002 (20 μM) or Akt inhibitor (20 μM) for 30 min prior to Aβ treatment for 24 h. Cells were blotted with p-mTOR (Ser 2448), mTOR, p-Raptor (Ser 792), Raptor and β-actin specific antibodies. n = 4. Each western blot image was presented as representative image. Quantitative blot data are presented as a mean ± SE. n = 3. *p < 0.05 vs. control, #p < 0.05 vs. Aβ treatment. (E,F) SK-N-MC cells and mouse hippocampal neurons were immunostatined with p-mTOR (Ser 2448) specific antibodies, and visualized by confocal microscopy. Scale bars, 100 μm (magnification, ×400). Data are presented as a mean ± SE. n = 3.