1. Floating human frontal cortex sections of 40 μm thickness were obtained from Banner Sun Health Research Institute in Sun City, Arizona (4 control and 3 LRRK2 G2019S mutation carrier subjects).
- None of the control subjects had a history of dementia or neurological or psychiatric disorders at the time of death (See Supplemental Table 1). Informed and written consent was obtained from the donors.
2. For immunostaining, sections were washed in 1× TBS containing 0.3% Triton X-100 (TBST), blocked in 3% NGS diluted in TBST, and incubated in primary antibody 2-3 nights at 4°C with shaking.
3. Primary antibodies used were GFAP (chicken, 1:250; AB5541, Millipore Sigma) and phospho-ERM (Rabbit, 1:250; #3726, Cell Signaling).
4. Following primary incubation, sections were washed in TBST, incubated in Alexa Fluor conjugated secondary antibodies diluted 1:200 (Life Technologies) for 2-3 hours at room temperature, washed with TBST, and mounted onto glass slides using a homemade mounting media (90% Glycerol, 20 mM Tris pH 8.0, 0.5% n-Propyl gallate) and sealed with nail polish.