Rabbit anti neun

Rabbit anti-Neuroligin-1 and anti-Neuroligin-3 were previously described53 (link). Rabbit anti-GluR2/3 was a kind gift from Dr.Nathalie Sans. The following commercially available antibodies were used: rabbit anti-vesicular glutamate transporter 1 (vGluT1), rabbit anti-vesicular GABA transporter (vGAT), mouse anti-Vesicle associated membrane protein 2 (VAMP2), rabbit anti-Homer1, mouse anti-gephyrin, mouse anti-NR1 (Synaptic Systems), rabbit anti-GAPDH (Enogene), mouse anti-actin (Sigma), sheep anti-parvalbumin (PV) (R&D), goat anti-calretinin (CR) (Swant), mouse anti-GAD67, anti-NR2A (Upstate Biotechnology), mouse anti-NR2B, mouse anti-PSD95, mouse anti-GABAβ3, mouse anti-Kv3.1b (Neuromab), mouse anti-GAD65 (Developmental Studies Hybridoma Bank), mouse anti-MAP2 (Chemicon), rabbit anti-NeuN (Novus Biologicals), mouse anti-CaMKII (ABR Affinity BioReagents), rabbit anti-acetylhistone3 (Cell Signalling).
Secondary antibodies with minimal interspecies cross-reactivity conjugated to cyanine and Alexa 633, 546 or 488 dyes (Jackson ImmunoResearch and Invitrogen) were used for visualization in immunostaining. Secondary HRP conjugated anti-mouse and anti-rabbit IgG, and IRDye 680 coupled anti-Mouse and IRDye 800 coupled anti-Rabbit IgG for quantitative western blotting were purchased from Jackson and LI-COR Biosciences, respectively.

The 10 μm thick sections were collected on poly‐L‐lysine‐coated glass slides and treated for hematoxylin–eosin (HE) staining and immunofluorescence analysis. For immunofluorescence analysis, sections were first blocked in 10% goat serum in PBST (0.01 M PB containing 0.05% v/v Tween 20) for 1 h at RT and then treated with rabbit anti‐NeuN (1:500; Novus), mouse anti‐GFAP (1:1000; Proteintech), Iba1 (1:500; Wako) and CD206 (1:1000; Proteintech) antibodies overnight at 4°C followed by incubation with Alexa Fluor™ 594‐conjugated goat anti‐rabbit IgG (Invitrogen) and Alexa Fluor™ 488‐conjugated goat anti‐mouse IgG (Invitrogen). The nuclei of the total cells were indicated by DAPI staining. Fluorescence signals were visualized under a fluorescence microscope (Zeiss Axio Imager. 2). GFAP represented astrocytes, NeuN represented neurons, Iba1 represented microglia, and CD206 represented M2‐type cells.