Ab16046

Adult mice (P42 or P56) underwent cardiac perfusion using 1% heparin and 4% paraformaldehyde. Dissected brains were stored in 4% paraformaldehyde overnight. Coronal sections (50 μm) were prepared using a vibratome (Leica). Sections were permeabilized with 1× phosphate-buffered saline (PBS) with 0.5% Triton X-100, blocked with 5% serum (goat/donkey), and incubated with primary antibodies (NeuN, Milipore, MAB377, 1:500; Cux1, Santa Curz, sc-13024, 1:500; FoxP2, Abcam, ab16046, 1:500; parvalbumin, Millipore, MAB1572, 1:1000) and Alexa Fluor 594 AffiniPure Donkey Anti-ms IgG (H+L) (Jackson, Cat # 715-585-150, 1:1000), Alexa Fluor 594 AffiniPure Donkey Anti-rb IgG (H+L) (Jackson, Cat # 711-585-152, 1:1000), and Alexa Fluor 488 AffiniPure Donkey Anti-ms IgG (H+L) (Jackson, Cat # 715-545-151, 1:1000). For Nissl staining for gross morphological analysis, brain slices were incubated with diluted Nissl (ThermoFisher; 1:1000) for 30 min. The shapes of brain structures were marked manually, and each of area was measured in a blind manner. Images were acquired using LSM 780 (Carl Zeiss) or AxioScan.Z1 (Carl Zeiss).