Protein samples were prepared from freshly dissected brain and used for semi-quantitative western blot analysis as previously described [33 (link)]. Primary antibodies were used in following conditions: anti-histone H3 (1:2,000; Abcam, Cambridge, MA), anti-acetyl histone H3 (1:2,000; Abcam, Cambridge, MA), anti-β-actin (1:10,000; Abcam, Cambridge, MA), anti-HDAC1 (1:2,000; Cell Signaling Technology, Danvers, MA), anti-HDAC2 (1:2,000; Cell Signaling Technology, Danvers, MA), anti-HDAC3 (1:2,000; Cell Signaling Technology, Danvers, MA), anti-iba1 (1:2,000; Wako, Osaka, Japan). Properly matched secondary antibodies were used before ECL reaction: anti-mouse IgG (1:2,000; Vector Laboratories, Burlingame, CA), anti-rabbit IgG (1:10,000; Vector Laboratories, Burlingame, CA).
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Western Blot Analysis of Histone Modifications and HDAC Proteins
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- Primary antibodies used at various concentrations: anti-histone H3 (1:2,000), anti-acetyl histone H3 (1:2,000), anti-β-actin (1:10,000), anti-HDAC1 (1:2,000), anti-HDAC2 (1:2,000), anti-HDAC3 (1:2,000), anti-iba1 (1:2,000)
- Semi-quantitative western blot analysis of protein samples prepared from freshly dissected brain
- Properly matched secondary antibodies used before ECL reaction: anti-mouse IgG (1:2,000), anti-rabbit IgG (1:10,000)
- Positive control: None specified
- Negative control: None specified
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