Nuclear Protein Extraction and EMSA Analysis

Nuclear proteins were extracted from the cells and dissected cortical tissues using a commercially available nuclear extraction kit and an EMSA kit (Panomics, Fremont, CA, USA). As with our previous studies [31 (link)], nuclear extracts (5 μg) were reacted with AP-1 oligonucleotide (5′-CGCTTGATGAGTCAGCCGGAA), NF-κB oligonucleotide (5′-AGTTGAGGGGACTTTCCCAGGC), Stat oligonucleotide (5′-ATCGTTCATTTCCCGTAAATCCCTA), or CREB oligonucleotide (5′-AGAGATTGCCTGACGTCAGAGAGCTAG). The DNA/protein complexes in the membranes were visualized by enhanced chemiluminescence Western blotting reagents and quantitated using a computer image analysis system (IS1000; Alpha Innotech Corporation).