Ab1557p

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.

Protein lysates from cortical neurons, 3xTg-AD mice and human brains were separated by SDS-PAGE using 7.5% Tris-Glycine polyacrylamide gels (Bio-Rad). Electrophoresis was conducted in a Tris-Glycine buffer (25 mM Tris, pH 8.3, 192 mM glycine, 0.1% SDS in dH₂O) by using the Criterion cell system (Bio-Rad). Blots were developed with rabbit anti-NR2B (1:1000, Millipore, #AB1557P), rabbit anti-pSer¹³⁰³NR2B (1:1000, Millipore, #07-398), rabbit anti-NR2A (1:1000, Millipore, #AB1555P), rabbit anti-PSD95 (1:500, abcam, #ab18258), mouse anti-synaptophysin (1:500, Millipore, #MAB329), rabbit antiphosho PKC (1:1000, Cell Signaling, #9371), rabbit anti-PKC (1:1000, abcam, #ab179521) and rabbit anti-β-actin (1:5000; Sigma-Aldrich, #A2066). Secondary antibodies conjugated with horseradish peroxidase (HRP) were purchased from Sigma (1:5000, sheep antimouse-HRP #A6782, goat antirabbit-#HRP A6154).