Gel electrophoresis and Western blotting were as described (e.g., Peng et al. 2011 (link)) using rabbit polyclonal anti-phospho-Ser845-GluA1 (1:1,500; AB5849, Millipore, Temecula, CA), mouse monoclonal anti-GluA1 (1:1,500; MAB2263, Millipore), rabbit polyclonal anti-GluA2 (1:1,000; PA1-4659, Thermo Scientific, Rockford, IL), rabbit polyclonal anti-PSD95 (1:1,000; AB9708, Millipore), mouse monoclonal anti-GluA3 (1:500; MAB5416, Millipore), and mouse monoclonal anti-α-tubulin (1:5,000; T6199, Sigma-Aldrich, St. Louis, MO). Immunoblots were analyzed using NIH Image J software. Following densitometry, intensities of bands corresponding to GluA1, GluA2, GluA3, and p-Ser845-GluA1 for each sample were divided by intensities of the corresponding α-tubulin bands. Results were expressed in comparison to the normalized control, defined as the AL group that received the control treatment. Results were analyzed by two-way ANOVA, with significant interaction effects followed by comparison of cell means of interest using the error term from the ANOVA in the denominator of a t-statistic.
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Quantifying AMPA Receptor Subunits
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Variable analysis
independent variables
- Treatment
- Intensity of bands corresponding to GluA1, GluA2, GluA3, and p-Ser845-GluA1 for each sample
- Intensity of bands corresponding to α-tubulin for each sample
- Positive control: AL group that received the control treatment
- Negative controls: Not explicitly mentioned
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