Fig. 2

CO₂ inhalation during retrieval promotes GluR2 ubiquitination and degradation. a Schematic of experimental procedure of aversive conditioning (Av. C.), memory retrieval (Ret) and CO₂ inhalation, as described previously. Thirty minutes after, brain slices were prepared using a brain matrix, and amygdala isolated by a biopsy punch. Synaptosomal proteins were collected by using Syn-PER Reagent. Whole tissue lysates or synaptosomal proteins were subjected to SDS-PAGE and analyzed by Western blot. b Synaptosomal proteins collected from animals receiving aversive conditioning with or without Ret and with or without CO₂ are indicated. GluR2 or actin was detected by using anti-GluR2 or anti-actin antibody respectively. c GluR2 in whole tissue lysates collected from animals subjected to aversive conditioning and Ret/CO₂ was first concentrated by IP using protein A/G agarose in conjunction with monoclonal anti-GluR2 antibody. Ubiquitinated GluR2 was assessed by Western blot analysis. Some animals were given β-Lac after behavior treatment. The negative control was prepared using nonspecific mouse IgG. Data are representative of three independent experiments