Rabbit anti rest

Western blot analysis was carried out as aforementioned (Mateos et al. 2009 (link)). Following the transfer to a nitrocellulose membrane (Schleicher & Schuell, Germany), milk blocked blots were incubated with the following antibodies: mouse anti-PSD95 (1:1000; Abcam), rabbit anti-REST (1:1000; Millipore), rabbit anti-PTBP1 (1:1000; Bethyl Laboratories), mouse antipan SHANK (1:1000; Millipore), mouse anti-SNAP25 (1:1000; Transduction laboratories), rabbit anti-NeuN (1:500; Millipore) and rabbit anti-GAPDH (1:1000; Enzo) overnight. Secondary incubation was done using antirabbit or antimouse IgG at a 1:5000 dilution at room temperature (Amersham Biosciences, Little Chalfont, England). Immunoreactivity was detected by ECL detection system (Amersham, Biosciences, Little Chalfont, England). The densitometry analyses of the immunoreactive bands were performed using ImageJ software (NIH, MA, USA). Some immunoblots were stripped using Restore™ Western Blot Stripping buffer (Pierce, Rockford, IL, USA) at room temperature for 15 min and then blocked and re-blotted with other antibodies.

The antibodies for Western blot analysis used were: mouse anti-α-tubulin (1:10,000; Sigma, clone B512), rat anti-α-synuclein (1:100; clone 15G7), mouse anti-α-synuclein (1:1000; BD, Synuclein-1), rabbit anti-GFP (1:1000; Santa Cruz), rabbit anti-H3 (1:16000; abcam), rabbit anti-H3K4me3 (1:16000; Abcam), rabbit anti-H3K9me1 (1:16000; Abcam), rabbit anti-H3K9me2 (1:4000; CST), rabbit anti-H3K9me3 (1:16000; Abcam), rabbit anti-H3K27me3 (1:8000; Millipore), rabbit anti-H3K9ac (1:8000; Millipore), rabbit anti-H3K14ac (1:8000; EpiGentek), rabbit anti-H4 (1:16000; Abcam), rabbit anti-H4K12Ac (1:16000; Abcam), rabbit anti-HP1α (1:1000; CST), rabbit anti-EHMT2 (1:2000; abcam), rabbit anti-p38 (1:1000; CST), goat anti-REST (1:250; Santa Cruz) and goat anti-SNAP25 (1:1000; Santa Cruz). The HRP-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (1:10,000). The antibodies for chromatin immunoprecipitation (ChIP) used were: rabbit anti-EHMT2 (10 μg; Millipore), mouse anti-H3K9me2 (4 μg; Abcam), and rabbit anti-REST (10 μg; Millipore).