Map2 chicken

Cells were fixed with 4% PFA, permeabilized and blocked with 0.3% Triton X-100 and 3% Donkey Serum in TBS. Samples were incubated with the following primary antibodies overnight at 4 °C: MAP2 (Sigma, St. Louis, MO, USA, M1406,1:250); GFAP (Dako, Carpinteria, CA, USA, Z0334, 1:500); GAD65/67 (Abcam, Branford, CT, USA, ab11070, 1:500); KGA (Abcam, Branford, CT, USA, ab156876, 1:200); TUJ1 (Sigma, St. Louis, MO, USA, T8660, 1:400); MAP2 Chicken (Abcam, Branford, CT, USA, ab5392, 1:10000). Alexa Fluor Dye secondary antibodies were applied (1:500) at room temperature for 30 min. Images were acquired with a spectral confocal microscope LSM710 (Zeiss, Matesalka, Hungary).

Cortical neurons (DIV14) were fixed with 4% PFA for 30 min at room temperature, permeabilized with 0.05% Triton and blocked with 10% normal donkey serum in PBS. Cells were stained overnight at 4°C with antibodies directed against PGC-1α (PGC-1α, rabbit, 1:200; Novus Biologicals; RRID: AB_1522118) and microtubule-associated protein 2 (MAP2, chicken, 1:2500; Abcam; RRID: AB_2138153). As secondary antibodies we used Alexa Fluor 488-coupled donkey antibodies recognizing chicken IgG (1:800, Jackson; RRID: AB_2340375), Cy3-coupled donkey antibodies recognizing mouse IgG (1:800, Jackson; RRID: AB_2340816), and Alexa Fluor 647-coupled donkey antibodies recognizing rabbit IgG (1:800, Abcam; RRID: AB_2752244). Images were taken with a Zeiss LSM 700 confocal microscope.