Anti eaat1

Total proteins were extracted through a conventional method using RIPA lysis buffer (Beyotime Biotechnology, China) with protease inhibitor cocktail (Beyotime Biotechnology, China) at 4 °C. Protein concentrations were determined by BCA protein assay (Beyotime Biotechnology, China), and equal amounts of total proteins were separated by 10% SDS-PAGE (Thermo Fisher Scientific, USA), and then transferred onto PVDF membranes (Millipore, USA). Next, the membranes were blocked with 5% nonfat milk in 0.1% Tween washing buffer for 1 h at room temperature, and then incubated with primary antibodies including anti-GLS (Abcam, USA), anti-GRIA3 (Abcam, USA), anti-GRIN1 (Novus, USA), anti-EAAT1 (Abcam, USA) and anti-GAPDH (Abcam, USA) overnight at 4 °C. On the next day, the membranes were incubated with the conjugated secondary antibody (Invitrogen, USA) for 1 h at room temperature and the bands were visualized using ChemiDoc Touch Gel Imaging System (Bio-Rad, USA). The blots were quantified by densitometry using ImageJ software (NIH).