Journal: PLoS ONE

Article Title: Dietary Restriction Affects Neuronal Response Property and GABA Synthesis in the Primary Visual Cortex

doi: 10.1371/journal.pone.0149004

Figure Lengend Snippet: The top panel shows a typical sample of a Western blot result from each DR cat (DR1-4) and control cat (NC1-4) using GAD67 and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) antibodies. The bottom panel shows the average optical density (OD) of GAD67 bands normalized against the corresponding GAPDH in both groups of cats. The average normalized optical density of GAD67 in DR cats was significantly higher than that in the control (** p<0.01).

Article Snippet: The membrane was blocked with 5% non-fat dry milk in TBS-Tween 20 for one hour and incubated overnight at 4°C in TBS-Tween 20 containing a rabbit polyclonal antibody GAD67 (dilution 1:200; Boster Bio-engineering Limited Company) or a rabbit polyclonal antibody GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (dilution 1:5,000; Sangon Biotechnology).

Techniques: Western Blot, Control

Journal: EBioMedicine

Article Title: Inhibition of Nwd1 activity attenuates neuronal hyperexcitability and GluN2B phosphorylation in the hippocampus

doi: 10.1016/j.ebiom.2019.08.050

Figure Lengend Snippet: Nwd1 location in excitatory synapses acute seizures brain tissues. (a–b) Immunofluorescence staining for Nwd1 in the cerebral cortex of individuals with TLE patients. Nwd1 co-localizes with excitatory synapses marker postsynaptic density protein PSD95 but presynaptic vesicular marker Vglut1 (a) and less in inhibitory synapses marker GAD67 (b). (c–d) Immunofluorescence of Nwd1 in the hippocampi from KA-induced acute seizures mice show that Nwd1 colocalizes with PSD95 and lacks of colocalization of Nwd1with Vglut1 (c) and less in GAD67 (d) in the neuronal cells.

Article Snippet: The following primary antibodies were used in this study: a rabbit anti-NWD1 pAb (1:500, Proteintech Group, Inc., Cat# 25025–1-AP); a rabbit anti-GAPDH mAb (1:1000, Proteintech Group, Inc., Cat# 10494–1-AP); a guinea pig anti-MAP2 mAb (1:300, Synaptic Systems, Cat# 188004); a mouse anti-GFAP mAb (1:200, Proteintech Group, Inc., Cat# 60190–1-Ig); a chicken anti-GAD67 mAb (1:300; Synaptic Systems, Cat# 198006); a goat anti-PSD95 mAb (1:300, Abcam, Cat# ab12093); a mouse anti-PSD95 mAb (1:300, Abcam, Cat# ab13552); a guinea pig anti-Vglut1 mAb (1:300; Synaptic Systems, Cat# 135304); DAPI (1:50, Sigma-Aldrich, Cat# D9542); a rabbit anti-GluN2A pAb (1:500, Proteintech Group, Inc., Cat# 19953-1-AP); a mouse anti-GluN2B mAb (1:500, Abcam, Cat# ab28373); a rabbit anti-GluN2B pAb (1:500, Abcam, Cat# ab65783); a rabbit anti-GluN2B-Tyr1472 pAb (1:500, Abcam, Cat# ab3856,); and a rabbit anti-GluN2B-Ser1480 pAb (1:200, Bioss, Cat# bs-5382R); a mouse anti-Beta Actin Antibody mAb (1:1000, Proteintech Group, Inc., Cat# 60008-1-Ig).

Techniques: Immunofluorescence, Staining, Marker