Fig. 9

ApoE4 and Aβ42 induced cPLA2 activation in human postmortem frontal cortical synaptosomes. a, Synaptosomes were incubated in oxygenated Kreb’s Ringer with different reagents: rApoE3, rApE4, Aβ42, Aβ42/rApoE4 for 30 min, followed by 15 min incubation with Kreb’s-Ringer (control), TNFα/IFNγ or ceramide-1-phosphate, respectively. The synaptosomes were harvested and homogenized, and the membrane and cytosolic fractions of synaptosomes were isolated by centrifugation. cPLA2 was enriched in both fractions by immunoprecipitation with anti-cPLA2 antibodies, and the p-cPLA2 and total cPLA2 were detected by western blot. b-d, Densitometric quantification of blotting shown in A