After washing the secondary antibodies with PBS 3 times at ten-min intervals, coverslips were being mounted on to glass slides employing the Vectashield mounting medium (Vector Laboratories), and examined under a laser confocal fluorescence microscope (LSM-710, Carl Zeiss)
Brain tissues were fastened in four% paraformaldehyde in .one M neutral phosphate buffer, adopted by cryoprotection in thirty% sucrose answer overnight, and then 10 mm sections were being well prepared…