Mouse anti mag

The protein concentration of each sample was measured using the Lowry method and the colorimetric reaction was measured on a SpectraMax M3 plate reader (Molecular Devices). 10–40µg of total protein was loaded on mini Protean TGX Precast gels (Biorad). After electrophoresis, proteins were blotted on a PVDF membrane (Biorad) and blocked in 5% Blotting Grade Blotter (Biorad) diluted in Tris buffered saline (TBS) containing 0.1% Tween 20 (TBST). Blots were then incubated with either of the following antibodies: chicken anti-human MCT1 antibody, mouse anti-PLP (MilliporeSigma), mouse anti CNPase (MilliporeSigma), rabbit anti-MCT2 (MilliporeSigma), rabbit anti-GLUT1 (abcam), rabbit anti-GLUT3 (abcam), mouse anti-MBP (Biolegend), mouse anti-MOG (MilliporeSigma), rabbit anti-Connexin-43 (MilliporeSigma), mouse anti-GAPDH (Thermo Scientific), and mouse anti-MAG (MilliporeSigma)(for more details see Key Resources Table). Membranes were overnight incubated with primary antibodies at 4°C. Membranes were washed with TBST and incubated with HRP labeled secondary antibodies for 1 hour at room temperature. After two washes in TBST and one wash in TBS, membranes were exposed to ECL reagent for 2 min. (Thermo Scientific) and chemiluminescent signal was detected with the LAS Imager 4000 (GE Healthcare).