Figure 2From: Apolipoprotein E4 (1–272) fragment is associated with mitochondrial proteins and affects mitochondrial function in neuronal cellsApoE4 interacts with the subunits of mitochondrial respiratory complex III in Neuro2a cells. Neuro2a cells were cotransfected with mammalian expression plasmids encoding FLAG-apoE4(1–272) or FLAG-apoE4(1–299) and plasmids encoding mouse UQCRC2, human HA-UQCRC2, or human HA-cytochrome C1, all of which are candidate proteins suggested to be associated with apoE4 (Table 1). Twenty-four hours following the transfection, the cells were harvested and treated with 500 μl of Triton X-100 solubilization buffer to obtain cell lysate. The cell lysate was then incubated with anti-FLAG M2-agarose affinity resin, and the protein binding to the affinity column was eluted using FLAG peptide, and the eluted protein was then analyzed by western blotting with anti-UQCRC2 (mouse UQCRC2) antibody (A), anti-HA antibody (human UQCRC2) (B), or anti-HA antibody (human cytochrome C1) (C).Back to article page