Tissue sections from human AD brain were treated with cold (+4°C) 99% formic acid for 1 minute at RT (25°C).
In our series, 5 out of 13 patients with an isolated relapse in the brain were treated with ICE chemotherapy, of which three patients achieved a long-term complete response.
Finally, to assess how the data suggesting that inhibition of LRRK2 induces autophagy in model cell lines relates to primary cells, primary astrocytes isolated from rat brain were treated with LRRK2-in1.
Upon photoactivation, the brain was treated with collagenase (2 mg/ml, Sigma) for 30 s, the glial sheath removed using fine forceps, and the brain was mounted again in Sylgard-coated Petri dish in saline.
After permeabilization with PBS containing 2% Triton X-100, the brains were treated with 70% formic acid (Sigma), and stained with a mouse monoclonal anti-Aβ antibody (Chemicon) followed by detection with biotin-XX goat anti-mouse IgG and streptavidin-Oregon Green 488 conjugate (Molecular Probes).
Paraffin sections of AD brains were treated with 10 μg/mL Proteinase K (Pro-K) for 30 min after autoclaving (Ac) and formic acid (FA) treatment.
To confirm these findings biochemically, Sarkosyl-insoluble fractions from two AD brains were treated with trypsin or Pro-K, then immunoblotted with RD3, RD4, anti-4R and anti-pS396 (Fig. 1g j).
Free-floating sections (50-μm thickness) from these brains were treated with 2 N HCl in distilled water (30 min) and then with 0.1 M sodium tetraborate (5 min).
Patients with metastatic brain lesions were treated with 10 consecutive fractions of radiotherapy (whole brain, 3 Gy/fraction, day 1 12) followed by a booster dose of 9 Gy (3 Gy/fraction, day 21 23).
Briefly, PrPres was obtained after mouse brain homogenates were treated with proteinase K (Roche, Meylan, France) (10 µg/100 mg brain tissue for 1 h at 37°C) and concentration by ultra-centrifugation (100,000 g for 2 h on a 10% sucrose cushion).
