Figure 2.

Expression levels of the ATG12–ATG5 ubiquitin-like conjugation system and activity of the MTOR-dependent pathway remained unchanged in APP-PSEN1-SREBF2 mice. (A) mRNA levels of the indicated autophagy-related proteins in brains from 7-mo-old WT and mutant mice analyzed by qRT-PCR. Absolute mRNA values were determined, normalized to Rn18s ribosomal RNA, and reported as relative levels referred to the expression in WT mice. (B) Protein expression levels were assessed in brain homogenates from 7-mo-old WT and mutant mice. Shown are representative immunoblots for ATG12–ATG5 conjugate, ATG7, and ATG5. (C) Western blot analysis of AMPK-dependent phosphorylation of ULK1 at Ser317. Cellular lysates from SH-SY5Y cells starved in FBS-depleted medium for 24 h were used as positive control (SC). (D) Representative immunoblots showing the MTOR-dependent phosphorylation status of RPS6KB1. In western blot analyses, densitometric values of the bands representing the specific protein immunoreactivity were normalized to the values of the corresponding ACTB bands and expressed as relative intensity values. n=3.