Pull-down Assays

To pull down proteins from brain lysates and LP2(P4) fractions, 1-2 mg of the

sample was incubated with 25-50 μg of glutathione beads containing GST fusion

proteins for 12-16 hrs.

The mixtures were washed three times with 1X PBS with

0.15% Triton X-100, and then resuspended in 20 μl of 1X NuPAGE LDS sample

buffer for NuPAGE and immunoblotted analysis.

Alternatively, the samples were

resuspended in 50μl of PBS for proteomics studies and stored at -20 °C before

quality control and proteomics analysis.