Anti map2ab

Cells were fixed in 4% paraformaldehyde for 10 min and blocked for 1 h at room temperature (RT) with blocking buffer (5% FBS and 0.3% Triton X-100). After blocking, the cells were incubated with the following primary antibodies for 2 h at RT: anti-βIII-tubulin (1:500, Covance), anti-Map2ab (1:500, Sigma), anti-GFP (1:500, Aves), anti-NeuroD1 (1:200, Abcam), anti-Tmem119 (1:500, Abcam), anti-CD68 (1:500, Bio-Rad), and anti-Iba1 (1:500, Abcam). Stained cells were visualized with a fluorescence microscope (Axiovert 200 M, Zeiss) and a confocal microscope (LSM800, Zeiss). Fluorescence intensity of the cell soma was quantified using LAS AF (Leica) or ZEN (Zeiss).

Cells were washed with PBS and fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO) for 15 min, then permeabilized in PBS containing 0.3% (w/v) Triton X-100 for 10 min, lastly blocked in PBS containing 4% (w/v) BSA for 2 h. Cells were incubated with the following primary antibodies (1:250 dilution) for 2 h at room temperature): anti-βIII-tub (Biolegend, cat. 801201), anti-MAP2ab (Merck Millipore, cat. AB5622), anti-GFAP (Biolegend, cat. 83721), and anti-GABA (Sigma Aldrich, cat. A2052). Subsequently, the cells were rinsed three times with 0.1% (w/v) BSA in PBS-Tr and incubated with the secondary antibody (1:500 dilution) for 1 h at room temperature in the dark (Alexa Fluor 488 A11029, and Alexa Fluor 568, A11036, Invitrogen). Nuclei were counterstained with DAPI (Invitrogen, cat. 1:10000) for 45 min. Cell images were acquired using a Keyence fluorescence microscope BZ-X710E equipped with the BZ-X800 Analyzer software (Keyence, Osaka, Japan) with a 20X Plan-Apo Gamma NA 0.75 objective and fluorescence filter set for GFP, TRITC, and DAPI.