Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: FXR1P is a GSK3β substrate regulating mood and emotion processing

doi: 10.1073/pnas.1506491112

Figure Lengend Snippet: High doses of valproate modulate ERK2 activity through a βARR2-dependent mechanism. (A) Quantitative immunoblot analysis of phosphoprotein levels of DARPP32, CREB, JNK3, and ERK2 proteins in the striatum of C57Bl6J WT mice at different times following acute treatment with valproate (sodium valproate, 400 mg/kg i.p.), and dose–response inhibition of ERK2 phosphorylation/activity by valproate as measured by quantitative immunoblot analysis 120 min after drug injection. (B) Quantitative immunoblot analysis of pERK2 levels in the striatum and frontal cortex of WT mice after VAL 10 or VAL 25 chronic valproate treatment or vehicle. (C) Quantitative immunoblot analysis of pERK2 levels in the striatum and frontal cortex of βARR2-KO after VAL 10 or VAL 25 chronic valproate treatment or vehicle. For all analyses, respective total protein signal was used as the internal reference for phosphoproteins. Data (means ± SEM) were normalized to average protein levels in vehicle-treated animals from the same genotype. *P ≤ 0.05. Student double-tailed t test was performed (drug vs. vehicle within each genotype). n = 5 mice per group. Detection and quantification of immunoblot signal were performed within a linear signal range using near-infrared fluorescence and a LiCor Odyssey.

Article Snippet: The anti–β-catenin, anti–phospho-GSK3β (Ser9), anti–phospho-CREB, anti-CREB, anti–phospho-JNK3, anti-JNK3, anti-p44/42, anti–phospho-p44/42 ERK, and anti-DARPP32 mouse monoclonal antibodies were purchased from Cell Signaling Technology.

Techniques: Activity Assay, Western Blot, Inhibition, Phospho-proteomics, Injection, Fluorescence