Computational protocol: Antidepressant Effects of Abscisic Acid Mediated by the Downregulation of Corticotrophin-Releasing Hormone Gene Expression in Rats

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Protocol publication

[…] The protein of the BE(2)-C cells was extracted using whole cell lysates via a radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail (Roche). The protein samples were separated on a 12%-SDS-polyacrylamide gel electrophoresis plate and detected using antibodies against RARα (ab28767, Abcam), RARβ (sc-552, Santa Cruz), and either β-actin-HRP (KC-5A08, KangChen) or GAPDH-HRP (KC-5G5, KangChen) as an internal control. These samples were then processed with an appropriate HRP-conjugated secondary antibody with regard to the protein of interest. The protein levels were analyzed using ImageJ (Wayne Rasband) and normalized relative to the internal control. [...] Data analyses were performed using SPSS Statistics 19.0 (IBM). The values are expressed as the means ± SEM. The differences between the two groups (e.g., the protein levels of RARα with or without ABA treatment) were tested using student’s t-test. The differences between the three groups or above (e.g., the differences in the ABA concentrations between the different rat brain regions) were tested using one-way analyses of variance (ANOVAs), followed by a LSD post-hoc test. P-values less than 0.05 were considered significant. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Rattus norvegicus
Diseases Amino Acid Metabolism, Inborn Errors, Hypothalamic Neoplasms
Chemicals Abscisic Acid, Corticosterone, Sucrose