The mouse spinal cord tissue was fixed with 4% paraformaldehyde for 24 hours, and cross-sections of the spinal cord tissue were obtained after being embedded in paraffin. Anti-GluR2-p was used to detect corresponding proteins in the sections. The sections were stained with diaminobenzidine for 15 seconds.
Donkey serum (5%) was used to block the nonspecific binding sites at room temperature for 1 hour. The sections were incubated with primary antibodies against PICK1 and ICA69 overnight at 4°C. After washing with phosphate-buffered saline thrice, the sections were incubated with the fluorescent secondary antibodies (Jackson) DyLight 488 goat anti-mouse lgG (H + L) and DyLight 594 donkey anti-rabbit lgG (H + L) for 1 hour at room temperature. Finally, the nuclei were stained with 4',6-diamidino-2-phenylindole and imaged with Leica Sp5 II laser confocal microscope.
Donkey serum (5%) was used to block the nonspecific binding sites at room temperature for 1 hour. The sections were incubated with primary antibodies against PICK1 and ICA69 overnight at 4°C. After washing with phosphate-buffered saline thrice, the sections were incubated with the fluorescent secondary antibodies (Jackson) DyLight 488 goat anti-mouse lgG (H + L) and DyLight 594 donkey anti-rabbit lgG (H + L) for 1 hour at room temperature. Finally, the nuclei were stained with 4',6-diamidino-2-phenylindole and imaged with Leica Sp5 II laser confocal microscope.