Mitochondrial Mislocalization Underlies Aβ42-Induced Neuronal Dysfunction in a Drosophila Model of Alzheimer's Disease
Figure 6
Phosphorylation of dCREB is not reduced in head extracts from Aβ42 flies at 25 dae.
(A) Anti-RRxpS/T detects phosphorylation of Drosophila CREB (dCREB) at Ser231. dCREB Ser231, the site equivalent to Ser133 of mammalian CREB, is the only RRxpS/T site in dCREB. Head extracts were subjected to immunoprecipitation using anti-RRxpS/T, followed by Western blotting with anti-dCREB. The specificity of the antibody was confirmed using loss-of-function dCREB mutant flies (CREBS162). A low level of expression of a dCREB transgene was used to rescue lethality of CREBS162 (CREBS162+hs-dCREB) [59]. The signal detected by anti-dCREB was reduced in these flies. (B) Head extracts from control flies or from Aβ42 flies at 25 dae were subjected to immunoprecipitation using anti-RRxpS/T, followed by Western blotting with anti-dCREB. The phosphorylated CREB levels were normalized by the CREB level detected by Western blotting of the crude extract and are shown as ratios relative to controls. No difference in phosphorylated dCREB signal was detected (mean±SD, n = 4; p>0.05). (C) The total CREB level is not altered in Aβ42 fly brains. The CREB levels were normalized by the tubulin levels detected by Western blotting and are shown as ratios relative to controls. (mean±SD, n = 4; p>0.05).