phosphate buffered saline (PBS, pH 7.4) followed by 4% paraformaldehyde (pH 7.4 in 0.1M phosphate buffer). Brains were postfixed for 24 h in 4% paraformaldehyde then rinsed and stored in PBS until sectioning. Brains were sectioned coronally, at a thickness of 40 μm from Bregma 1.2 through −7.0, on a vibrating microtome (Leica VT1000S, Germany) and were collected and stored in cryoprotectant in 24 well plates at −20°C until use.
