Figure 2
From: Ubiquitin-proteasome dependent degradation of GABAAα1 in autism spectrum disorder
GABA A α1 levels are regulated through ubiquitin-proteasomal Degradation. (A) Primary cortical neurons (DIV 14) were treated with vehicle (control; DMSO), MG132 (20 μM), or lactacystin (50 μM) for 9 h, and lysates were subjected to proteasome activity assay. Results are mean ± SEM. *P <0.05 vs. control. (B-D) Primary cortical neurons (DIV 14) were treated with vehicle (control) or MG132 (20 μM), lactacystin (50 μM), or betulinic acid (20 μg/mL) for 9 h, and lysates were subjected to western blot analysis. The upper panel shows representative autoradiogram of GABAAα1 or GAPDH, and the lower panels represent fold change in normalized GABAAα1 protein levels in (B) MG132, (C) lactacystin, or (C) butelinic acid-treated neurons. Results are mean ± SEM. *P <0.05 vs. control. (E) Primary cortical neurons (DIV 14) were treated with vehicle (control) or MG132 (20 μM) for 9 h, and lysates were immunoprecipitated (IP) with an anti-GABAAα1 antibody, followed by western blotting (WB) with an anti- ubiquitin Lys48-specific antibody (lys48Ub) or anti-GABAAα1 antibody. IgG, IgG control. Results are representative of three independent experiments.