Ab124802

All chemicals were purchased from Sigma-Aldrich (St Louis, MO, USA) unless otherwise indicated. For western blots, primary rabbit antibodies against xCT, EAAC1, GLAST or GLT1 (ab37185, ab124802, ab416 or ab41621 respectively) were obtained from Abcam, Cambridge, MA, USA. Anti-LAT1 (sc-34554) from Santa Cruz Biotechnology, Santa Cruz, CA, USA. Rabbit anti-NR2A, anti-NR2B or mouse anti-GAPDH (AB1555P or AB1557P, MAB374 respectively) from Millipore, Bedford, MA, USA. Rabbit anti mouse-β-tubulin (T4026) from Sigma-Aldrich. Secondary goat anti-rabbit antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). For immunofluorescence staining chicken anti-MAP2 (ab5392) from Abcam. Anti-rabbit Alexa Fluor 594 (A11039) and anti-chicken Alexa 488 (A21207) secondary antibodies were obtained from Life Technologies, Carlsbad, CA, USA.

Cells were lysed in 1.5× Laemmli buffer, and protein concentrations were determined using BCA protein assay (ThermoFisher Scientific, Waltham, MA, USA). 20–40 μg of total protein extracts were separated by electrophoresis in 10% SDS-PAGE and transferred onto PVDF membranes (Immobilon, Merck Millipore Ltd., Tullagreen, Carrigtwohill, Co. Cork, Ireland). The membranes were blocked with 5% non-fat milk in PBS and incubated with anti-human primary antibodies against: xCT (1/1000; 12691; Cell Signaling Technology—CST, Leiden, Netherlands), phospho-GCN2 (1:500; ab75836; Abcam, Cambridge, UK), rabbit ATF4 (1:1000; 11815S; CST), phospho-S6RP (1:1000; 2215S, CST); phospho-S6K (1:1000; 9202S; CST), LAT1 (SLC7A5; 1:1000, KE026, TransGenic Inc., Illkirch-Graffenstaden, France), ASCT1 (SLC1A4; 1:1000; 84424; CST), ASCT2 (SLC1A5; 1:1000; 8057S, CST), EAAT3 (1:1000; ab124802, Abcam), EAAT4 (1:1000, ab186435, Abcam) or CHAC1 (1 μg/mL; AV42623; Sigma Aldrich). Protein loading control was verified by detection of tubulin (1:1000; MA5-16308; Thermo Scientific, Waltham, MA, USA) or ARD1 (rabbit anti-human/mouse; homemade [42 (link)]). Immunoreactive bands were detected with horseradish peroxidase antimouse or antirabbit antibodies (Promega, Medison, WI, USA) using ECL system (Merck Milipore, Watford, UK). Immunoblot analysis was performed using the LI-COR Odyssey Imaging System (Lincoln, NE, USA).

All chemicals were purchased from Sigma-Aldrich (St Louis, MO, USA) unless otherwise indicated. Methylarsonic acid (MMAV) disodium salt (99% pure) was obtained from Chem Service (West Chester, PA, USA). Sodium borohydride was obtained from EM Science (Gibbstown, NJ, USA). For Western blots, primary rabbit antibodies against xCT (ab37185), EAAT3 (ab124802), GLAST (ab416), GLT1 (ab41621), NR2B (ab65783), GluA2 (ab133477), Synaptophysin (SY38, ab8049), CBS (ab135626), and CSE (ab151769) were obtained from Abcam (Cambridge, MA, USA). Anti-LAT1 (sc-134994) and PSD95 (7E3, sc-32290) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-NR2A (AB1555P) and anti-GluA1 (AB1504) antibodies were purchased from Millipore (Bedford, MA, USA). Primary antibody against SLC1A4 (8442 s) and secondary goat anti-rabbit antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Secondary anti-mouse IgG antibody was purchased from Invitrogen (Waltham, MA, USA).