Pcmv6 xl vector

Cells were transfected with 100 nm of an AKT- or CREB-specific siRNA oligonucleotide. In the control experiments, the cells were transfected with 100 nm of scrambled siRNA. For ectopic expression of p65 or mPGES-1, the human A172 cells were transfected with 1.6 μg per dish of plasmid containing the p65 or mPGES-1 cDNA using lipofectamine 2000. In the control experiments, the cells were transfected with 1.6 μg per slide of the empty pCMV6-XL vector (Origene Technologies). The transfected cells were allowed to recover for at least 12 h in growth medium and then incubated overnight in serum-free medium before extraction.

Aβ₁₋₄₂ was synthesized by Qiangyao Biotechnology (Shanghai, China). BDNF was obtained from Sigma-Aldrich Corp (St. Louis, MO, USA). Antibodies specific for BDNF, PGC-1α, and FNDC5 were obtained from Abcam (Shanghai, China). Antibodies against Aβ (Stock #2454) and β-actin were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Of note, the Aβ antibody specifically reacts with human Aβ. PGC-1α or FNDC5 cDNA plasmids were obtained from Origene Technologies (Rockville, MD, USA) and subcloned into the pCMV6-XL vector. All reagents for the qRT-PCR and SDS-PAGE experiments were purchased from Bio-Rad Laboratories. All other reagents are from Invitrogen (Carlsbad, CA, USA) unless otherwise specified.